University of California, Los Angeles
Office for Protection of Research Subjects
HUMAN SUBJECT PROTECTION COMMITTEE (HSPC)
APPLICATION TO INVOLVE HUMAN SUBJECTS IN RESEARCH
INVESTIGATOR'S
ASSURANCE |
I certify that the information
provided in this application is complete and correct.
I understand that as
Principal Investigator, I have ultimate responsibility for the conduct of the
study, the ethical performance of the project, the protection of the rights and
welfare of human subjects, and strict adherence to any stipulations imposed by
the HSPC.
I agree to comply with
all UCLA policies and procedures, as well as with all applicable federal,
State, and local laws regarding the protection of human subjects in research,
including, but not limited to, the following:
�
performing the project
by qualified personnel according to the approved protocol,
�
implementing no changes
in the approved protocol or consent form without prior HSPC approval (except in
an emergency, if necessary to safeguard the well-being of human subjects),
�
obtaining the legally
effective informed consent from human subjects or their legally responsible
representative, and using only the currently approved, stamped consent form
with human subjects,
�
promptly reporting significant or untoward adverse
effects to the HSPC in writing within 5 working days of occurrence.
�
if I will be unavailable
to direct this research personally, as when on sabbatical leave or vacation, I
will arrange for a co-investigator to assume direct responsibility in my
absence.� Either this person is named as
a co-investigator in this application, or I will advise HSPC by letter, in
advance of such arrangements.
____________________________________________________________________
Principal Investigator���� �����������������������������������������Date
FACULTY SPONSOR�S
ASSURANCE |
By my signature as sponsor on this research
application, I certify that the student�
or� guest investigator is
knowledgeable about the regulations and policies governing research with human
subjects and has sufficient training and experience to conduct this particular
study in accord with the approved protocol.��
In addition,
�
I agree to meet with the
investigator on a regular basis to monitor study progress.
�
Should problems arise
during the course of the study, I agree to be available, personally, to
supervise the investigator in solving them.
�
I assure that the
investigator will promptly report significant or untoward adverse effects to
the HSPC in writing within 5 working days of occurrence.
�
If I will be
unavailable, as when on sabbatical leave or vacation, I will arrange for an
alternate faculty sponsor to assume responsibility during my absence, and I
will advise the HSPC by letter of such arrangements.
____________________________________________________________________
Faculty Sponsor *
(if PI is a student or a fellow)�������
Date
* The faculty sponsor must be a member of the UCLA
faculty.� The faculty member is
considered the responsible party for legal and ethical performance of the
project.
SECTION
II - FUNDING |
THIS SECTION
MUST BE COMPLETED
1. Check all of the appropriate
boxes for funding sources for this research, include pending funding source(s):
�X Extramural*����� UCLA Academic Senate����� Department�����
Gift����� Other:
* P.I. of Contract or Grant:� Paul Krogstad, M.D.����
Funding Source: National
Institutes of Health
2.�� If using an IDENTICAL protocol for more than one extramural funding proposal,
list all funding sources below.� Attach
an additional sheet if more space is needed.
a.�� P.I. of Contract or Grant:
Funding Source:
Contract or Grant No.:
Contract or Grant Title:
b.�� P.I. of Contract or Grant:
Funding Source:
Contract or Grant No.:
Contract or Grant Title:
3.�� STATEMENT OF FINANCIAL INTERESTS: If
you are required to submit either a Form 730-U* or a Form 740-U* to the Office
of Sponsored Research, please attach a copy of those form(s) with this
application.� See #9 of the Guidelines
for additional information regarding this requirement.
����� * Form
730-U, "Principal Investigator's Statement of Economic Interests" for
non-governmental funded projects
����� * Form
740-U, "Investigator's Statement of Financial Interests" for NSF or
PHS funded projects
4.�� Is this
application for the administrative approval for a training grant, a program
project, a multiple project grant, or a center grants?�� Yes��� X No���
If yes,see Guidelines #14.
If
this application is applying for an administrative approval for funding
purposes only and does not involve the participation of human subjects,
do
not complete the rest of this application.
SECTION
III - SUMMARY INFORMATION |
THIS SECTION
MUST BE COMPLETED
The review of research involving human subjects is
conducted by either the Medical Human Subject Protection Committee (MHSPC) or
the General Campus Human Subject Protection Committee (GCHSPC) depending on the
nature of the protocol.� The MHSPC is
composed of primarily medical specialists, and the GCHSPC has principally
socio-behavioral experts and some medical professionals.� To aid the OPRS staff in evaluating which
HSPC is most likely appropriate for the review of your protocol, please check
all appropriate boxes in this section.
1.���� Will you
perform medical procedures as part of this research proposal?�� XYes�
No
2.���� SUBJECT POPULATION:� (Check all appropriate boxes.)
X Children (see
Manual Chapters 4,6,8, & 10)���� Cognitively or psychologically impaired (see Manual Chapter 4)
Elderly (see
Manual� Chapters 4 & 10)������������ Institutional residents (see Manual Chapters 4 &
8)
Fetuses (see
Manual Chapter 8)������������������������ Human in vitro fertilization (see Manual Chapter 8)
Pregnant women (see
Manual Chapter 8)�������� Exclusion of minorities (see Manual Chapter 8)
Terminally ill (see
Manual Chapter 8)��������������� Prisoners or parolees (see manual Chapter8)
Comatose (see
Manual Chapter 4)�������������������� Non-English speaking (see Guidelines #11 & Manual Chapter 8)
Cancer patients (see
Guidelines #4)������������������ UCLA students/staff (see Guidelines #10 & Manual Chapter 8)
3.���� If the research involves any of the
following, check the appropriate boxes:
Interviews������������������������������������������������������������� X
HIV/AIDS
Survey/questionnaire������������������������������������������ X Clinical studies
Behavioral observation��������������������������������������� Investigational drugs (if checked, complete Section V)
Deception�������������������������������������������������������������� Investigational devices (if checked, complete Section VI)
Waiver of consent����������������������������������������������� Radiation (see
Guidelines #5)
Study of existing data (see Guidelines #12)��� Controlled substances (see Guidelines #6)
Study of human biological specimens�������������� Microorganisms or recombinant DNA (see Guidelines #7)
��� (see Guidelines #12)������������������������������������������� Potential development of commercial product from human
X Venipuncture (<450cc)����������������������������������������� ���� biological materials (see
Guidelines #8)
������� X
Genetic research���������������������������������������������������� X PI or Co-PI is the treating physician
4.���� LOCATION(S)
OF RESEARCH TO BE CONDUCTED AT:
������� X UCLA campus��������������������������������������������������������� Santa Monica-UCLA Medical Center
������� X
Other locations, specify: Children�s Hospital, Los Angeles
�������
5.���� LAY
LANGUAGE SUMMARY:� (Please use non-technical
language that is understood by nonscientific members to summarize the proposed
research project.� The information must
include: (1) a brief statement of the problem and related theory supporting the
intent of the study, and (2) a brief but specific description of the
procedure(s) involving the human subjects.�
Attach an additional page as necessary.�
However, please do not exceed one single-spaced, type-written page.)
The thymus is a
large gland found in the chest.� It is
the source of CD4+ and CD8+ T cells, which play a key role in the body�s
defense against infection and cancer.�
These cells are progressively lost due to HIV infection.� In people with HIV, treatment leads to an
increase in T cells.� It remains unclear
how much of the increase is due to production of new T cells by the thymus and
how much is due to the improved survival of existing T cells.�
The intent of
this study is to compare several aspects of the function of the thymus in
subjects 13-24 years old who acquired HIV at birth versus subjects who acquired
HIV through sexual activity or drug abuse versus subjects who are HIV
negative.� This will be done using blood
collections as well as a CT scan of the thymus.� In addition, a substudy will be performed during which subjects
will be admitted to the Clinical Research Center overnight.� They will be given either a sugar solution
or a water solution with a non-radioactive marker.� Blood will be collected so that this marker can be detected.
SECTION
IV - PROTOCOL SUMMARY |
THIS SECTION
MUST BE COMPLETED
INSTRUCTIONS: �In order to
review your proposal, the Human Subject Protection Committee (MHSPC) must have
all of the following information. Each topic must be titled using the boldface subheadings listed below.� State �Not Applicable� for topics that are
not applicable to your application.�
Address each topic independently in the sequence listed without reliance
on information covered under other subparts.�
Attaching sections of the grant application is not an acceptable
substitute.� Provide sufficient
information for effective review by all members of the HSPC, including
non-specialists.� Define all
abbreviations and terms not part of common language and use simple words and
sentence structure as much as possible.�
Unless justification is provided, Section IV of this application must
not exceed 10 pages (excluding references).�
Number each page, beginning with page one for the first page of Section
IV.
INFORMATION
REGARDING RENEWAL APPLICATION (1) |
1.���� Renewal Application:� What benefits to the participating subjects
or to the society have been derived?�
Please also provide a summary of the research activities during the
previous approval periods regarding the following issues:
a)���� How many
subjects have been enrolled since the date of last approval and since the
initial approval?
b)��� Has there
been any difficulty obtaining/retaining subjects or obtaining informed consent
during the previous approval period?� If
yes, describe:
����� Approximately
how many potential subjects have refused participation?
����� How many
subjects have voluntarily withdrawn participation at their own request?
����� How many
subjects have withdrawn participation at the request of the PI?
c)���� Have there
been any unexpected reactions or complications since last scheduled annual
review?�
If yes, please attach Adverse Event Reports (Form
HS-5).� If you have submitted the
Adverse Event Reports, please state so.
d)��� Approximately
how many more subjects are required to complete the study?
Not
applicable � not a renewal
PURPOSE OF
THE STUDY, THE BACKGROUND AND THE LITERATURE REVIEW (2-3) |
2.���� Purpose of the Study:� What are the specific scientific objectives
(aims) of the research?
1.
To compare quantitative parameters of thymopoiesis from adolescents/young
adults with perinatal HIV infection (PI-A) with those from age-matched
seronegative control subjects (SN-A), and youth with HIV infection acquired via
recent adult behaviors (AB-A).�
2.
To evaluate the impact of viral factors on thymopoiesis of HIV infected
adolescents.
3.
To examine the T cell receptor repertoire and CTL responses of
perinatally infected adolescents.
3. ��� Background: State the background of the
study.� Include a critical evaluation of
existing knowledge, and specifically identify the information gaps which the
project is intended to fill.� Describe
previous work in animal and/or human studies that provide a basis for the
proposed research and that support the expectation of obtaining useful results
without undue risk to human subjects.�
������� Note:�
Include appropriate citations to the scientific literature or attach a
copy of literature review.
Long term survival after
HIV perinatal infection.
Most cases of pediatric
HIV-1 infection result from perinatal infection, occurring either in utero or at the time of
delivery.� Other cases are acquired
postnatally via transfusion or breast-feeding.�
Regardless of the means by which infection is acquired, untreated
pediatric HIV infection is generally followed by the development of symptomatic
disease in the first year of life, and the development of AIDS in as many as
50% of children by 5 years of age [1,
2] .� In the
pre-HAART era, the median and mean survival times for perinatally infected
children were 8.0 and 9.4 years, respectively [2,
3] .� However,
survival into adolescence is now occurring in many cases. The number of these
individuals is unclear, but the most recent HIV/AIDS Surveillance Report [4] �notes 180
people who acquired HIV infection perinatally who have had AIDS diagnosed after
age 13.� It is likely that a much larger
number of children were diagnosed with AIDS prior to age 13, and are now
surviving into adolescence because of improvements in preventative care, and the
advent of highly active antiretroviral therapy (HAART).� In concert with the marked decrease in
perinatal transmission seen in the last seven years, this increasingly large
population of perinatally infected adolescents is changing the face of
Pediatric HIV/AIDS in the United States.
Survival into
adolescence is likely to be attributable to a combination of viral, host, and
treatment factors. Chemokine and chemokine receptor gene polymorphisms, HLA
type, and mutation of the viral nef
gene are potentially important variables [5]
.� However,
CCR5 deletions are uncommon in non-Caucasian populations that are most affected
by the HIV epidemic [5, 6] , and large deletions in nef appear to be rare.�
There have been two small reports [7,
8] �in which
mutations of the nef gene were found
in 8 long-term survivors of perinatal infection.� In these reports, two patients had large nef gene deletions, and had strikingly mild disease.� One was asymptomatic and on no antiretroviral
therapy at 10 years of age, and did not have detectable plasma HIV RNA.� The other was 12 years of age and had only
moderately symptomatic disease (CDC class B).�
Missense mutations and small deletions of nef were found in some of the others. In contrast, most long-term
surviving children have much more advanced disease than the subjects described
in these reports.� Nielsen et al [9] �reported a
multicenter study of pediatric long-term survivors, defined to be 8 or more
years of age.� Only 31/143 (21)% of
children with maternally derived HIV infection and 9/54 (17%) with transfusion
acquired infection had > 500 CD4+ T cells and no prior AIDS defining
conditions.� Thus, in most cases,
progressive immunodeficiency and AIDS are seen in adolescents who acquired HIV
infection perinatally or via transfusion in early childhood, suggesting that
these individuals are infected with fully pathogenic HIV variants.
Immunological changes
associated with HAART.
The accumulated experience with HAART in adults has shown us that the prolonged suppression of HIV viremia to �undetectable� levels will often (though not invariably) arrest disease progression and bring about significant immunological restoration.� Initially, the marked increase in CD4+ T cells seen with potent anti-retroviral agents was attributed to the reduction in virus-induced destruction or clearance of cells [10, 11] .� Later reports made it clear that redistribution of lymphocytes from lymphoid organs accounted for much of the initial increase in peripheral blood T lymphocytes [12-14] .� Subsequently, the qualitative and quantitative changes seen in response to HAART were better characterized.� A triphasic pattern of immune reconstitution was described by Autran et al. in studies of adults.� An early rise of memory CD4+ T cells was soon followed by improved T cell proliferation responses to recall antigens, and, finally, a late rise in putative na�ve CD4+ T cells (CD45RA+ CD62L+).� Functional improvements in cellular and humoral immune responses are often evident within months [15] . Similar patterns have been observed following HAART in children, although CD45RA+ T cells often increase in number during the first several months of therapy [16, 17] .� Immunological improvements are less well documented during HAART in children, but include improved responses to measles immunization and proliferation responses to Candida [18-20] .
These data and the demonstration of decreased risk of opportunistic infections following prolonged HAART therapy [21, 22] �have engendered optimism that suppression of HIV replication may be followed by at least partial restoration of normal immunity. However, it is clear that quantitative and functional immunological abnormalities generally persist.� T cell counts remain below the normal range in most adults, and opportunistic infections continue to occur in some individuals.� At a more subtle level, the T cell receptor repertoire continues to show marked perturbation even after long periods of successful HAART.� This has been assessed in a number of ways, but is perhaps most readily quantifiable by using RT-PCR to examine the size distribution of sequences in the third coding complementarity determining region 3 (CDR 3) of the T cell receptor (TCR) b chain. Insertion and removal of nucleotides during TCR recombination normally leads to a Gaussian distribution of the size of amplicons resulting from RT-PCR amplification of Vb mRNAs with primers that flank junction sites.� Skewed distributions in CDR3 size pattern detected by this �spectratyping� are seen in chronic HIV infection, and often persist despite successful HAART [23-25] .
Persistent
immunological defects may be more extreme following prolonged HIV infection in
childhood.� HIV infection in adults
occurs after immunological development is complete, and substantial
immunological experience with common pathogens has occurred.� Perinatal HIV infection in children is
characteristically is associated with high viral RNA levels throughout the
first several years of life, perhaps leading to a disruption of immunological
ontogeny, interfering with the acquisition of protective responses to CMV,
toxoplasmosis, and other microorganisms [26,
27] .� Moreover,
HIV has been associated with accelerated thymic involution, which could be
exacerbated by the sustained high-level viremia characteristic of early
childhood.� Indeed, an immunophenotype
consistent with DiGeorge anomaly has been described in children with rapid
progression of disease [28, 29] .� The
mechanism (s) underlying thymic injury by HIV are unclear, but the virus might
directly kill thymocytes, or disrupt the microenvironment of thymic dendritic
cells and epithelial cells required for normal thymopoiesis [30] . Although data from the SCID-hu mouse model indicate
that the thymic epithelial compartment may remain functional long after
depletion of thymocytes has occurred [31]
, there may be limits to this: age dependent
difference in responses to HAART have been noted.� Some studies indicate larger increases in CD4+ T cell counts are
seen in younger compared to older adults [32]
�and in
children compared to adults [33] .� HAART may be
less beneficial in perinatally infected adolescents, who have potentially had
the longest exposure to injurious effects of HIV replication on the
thymus.�
Patterns
and parameters of thymopoiesis.
�The potential impact of perinatal HIV
infection on the future survival of those who have survived into adolescence
must be considered in light of recent developments in the understanding of
thymopoiesis.
Until approximately
twenty years ago, the thymus was thought to enlarge in childhood, and then
involute during puberty under the influence of growth hormone and sex steroids.
More recent studies of anatomy and physiology of the normal thymus have called
this notion into question [30, 34] .� Careful
histological analysis of tissue revealed the thymus to be a chimeric organ
containing epithelial tissue, where thymopoiesis occurs, and non-epithelial,
perivascular space (PVS).�� In
childhood, the thymic epithelial space (TES) decreases progressively, while the
perivascular space gradually enlarges [35]
.� During
adolescence, the total amount of lymphoid tissue remains constant, owing to an
increase of the lymphoid PVS.� Following
adolescence, the PVS decreases, and fatty atrophy of the thymus is seen.
However, after accounting for the perivascular space, Steinmann demonstrated a
continuous loss of the lymphoid tissue in the cortex from 1 to 40 years of age [34] , without evidence of any change in the rate of loss
during or after puberty.� After the age
of 40, the rate of involution of the TES decreases, but loss of the tissue
involved in thymopoiesis continues. Still, the TES has been seen in
centenarians, and recent reports have provided evidence of functional thymic
activity in the fifth and sixth decades of life [36, 37] .� While the
thymus does not normally decrease in total volume with aging, HIV infection has
been associated with a decrease in thymic volume, and histological evidence of
premature atrophy, with decrease in TES, and an increase in the PVS.� This effect may be a direct effect of HIV
replication: HIV infected cells have been found in both the PVS and the TES [30] .� The recent
detection of infected na�ve peripheral CD4 T cells is also consistent with this
possibility [38, 39] .
If there is an effect of
puberty on the rate of involution of the thymus, it does not appear to be
readily detected in those not infected by HIV.�
However, delayed onset of puberty has been often seen in pediatric long-term
survivors of HIV infection [40] , and castration prevents post-pubertal thymic
involution in mice [41, 42] .� It is
conceivable that delayed puberty will have an effect on the rate of thymic
involution of perinatally infected adolescents.� If higher levels of testosterone and other androgens promote loss
of functional tissue, delayed puberty may actually be beneficial, from a
teleological perspective, in the face of ongoing HIV replication.
Until recently,
non-invasive measures of the amount of functional thymus did not exist. An
ideal laboratory method for the evaluation of HIV infected individuals would
quantify both current thymic activity and the amount of healthy tissue that
might persist.�
Computed tomography has
been used to examine the size of the thymus in adult lacking HHIV infections,
corroborating post-mortem studies that found no change in the size of the
thymus with age [43] .� Clearly,
radiographic imaging has significant limitations as a tool to evaluate the
capacity for thymus dependent regeneration of T cell populations during HAART
(or after cytotoxic chemotherapy or bone marrow transplantation).� However, McCune and his collaborators found
a correlation between thymic mass (any residual tissue not replaced by fat
scored as an ordinal variable on a five point scale) and CD4 T cell number in
HIV infected adults who were 20 to 59 years of age [44] .� They also
found a correlation between these estimates of the amount of thymic tissue, and
increased thymopoiesis during HAART [45]
.� Similarly,
Vigano et al used magnetic resonance imaging to monitor changes in thymic
volume during HAART in a cohort of children with a mean age of 9.8 years at
entry [46] .� The thymus
was markedly diminished in volume in HIV infected children with Class C
disease.� A rise in CD45RA+CD62L+ CD4+ T
cells during HAART and was statistically associated with a change in thymic
volume.� Thus, the size of the thymus,
as determined by radiographic imaging, has correlated with an increase in na�ve
T cells during HAART of both adults and children.
Until recently, no
methods existed to identify and quantify lymphocytes that have recently
emigrated from the thymus.� In chickens,
lymphocytes may be identified as recent thymic emigrants (RTE) with the
monoclonal antibody chT1 [47, 48] .� No
comparable cell surface marker exists for humans or other mammals, but methods
to quantify RTE have been developed based on the use of quantitative PCR
measurements of circular DNA molecules generated during excisional
rearrangement at the T cell receptor alpha [37,
49] �or beta [50] �chain loci.
The PCR assay described by Douek et al [37]
�is used to
quantify circular DNA molecules (termed signal joint TREC (sjTREC)) produced in
approximately 70% of the cells with excision of the d locus during maturation
of thymocytes into ab T cells [51-53]
.� In control
subjects, Douek et al found a nearly linear inverse correlation between age and
TREC number in CD4+ and CD8+ T cells.�
These findings are in accord with the histological studies described
above suggesting a gradual, progressive, diminution of the thymic epithelial
space throughout the first four decades of life, without evidence of marked
changes during puberty.�� In accord with
these data, we have also found that TREC values are very stable in serial
samples from HIV seronegative adolescents followed for up to 48 months (see
preliminary studies below).�
In agreement with
several studies from adults, decreased TREC numbers are seen in chronically HIV
infected children, and there appear to be age-dependent differences in
responses to HAART: the increase in TREC seen with HAART was greater in infants
than in children with a median age of nine years [54] .� Of note,
Hazenberg et al have recently called the utility of TREC quantitation as a
parameter of thymic output into question, noting that proliferation of na�ve T
cell populations may explain the drop in TREC seen after HIV infection has
occurred [55] .� However,
this interpretation been called into question, as it is based on the
measurement of TREC in measurement of cells identified as �na�ve� by expression
of CD27 and lacking CD45RO.� As noted by
Grossman and Paul, these markers would also be present in activated cells in
transition to the expression of markers identifying them as memory T cells.� For now, measurement of TREC appears to be a
useful parameter of thymic output [56]
.
The survival of T cells produced during
HAART could conceivably be influenced by the duration of previously
uncontrolled HIV infection.� Recent data
suggest that maintenance of na�ve and memory cells requires a supportive
environment in the periphery, which may be distinct for each population [57] .� While the
signals necessary for the maintenance of the na�ve T cells are still being
defined, current data suggest that na�ve CD4+ T cells are maintained without
proliferation by contact with cells bearing MHC class II molecules [58] .� Presumably,
this represents the peripheral lymphoid mass, implying that HAART may need to
bring about substantial normalization of lymph node architecture before
immunological reconstitution is optimal.�
The chances for reconstruction of this microenvironment may be lower
with longer periods of uncontrolled HIV replication after perinatal infection.�
Directly
measuring T cell dynamics in vivo may make it possible to determine if
differences in T cell survival play a role in the clinical heterogeneity of
long-term survivors of perinatal HIV infection. Hellerstein, McCune and their
collaborators have shown that this is possible, by developing methods to
determine rates of T cell turnover and cellular half-life [45, 59] . They administered prolonged infusions of glucose
labeled with deuterium, a non-�radioactive stable isotope, to uninfected and
HIV-1 infected adults.� Using gas
chromatography-mass spectrometry, the fractional replacement of deuterium for
hydrogen atoms was measured, allowing them to calculate the rate of turnover of
T cells in the peripheral blood, and to estimate the half-lives of CD4 and CD8
T cells.� In untreated HIV infected
subjects, T cells had decreased half-lives, and no compensatory increase in T
cell production. HIV treatment was associated with an increase in the absolute
production rate of peripheral blood T cells, and normal half lives and production
rates were restored after 12 to 36 months of HAART.� In preliminary experiments performed with the assistance of Dr.
Hellerstein, we have shown that these methods can be successfully applied to
newborn macaques weighing less than 1
kilogram.
��������������� Deeks et al, have also recently
helped explain the phenomenon of so-called �discordant responses� to HAART, in
which higher
CD4 T cell counts and peripheral blood TREC concentrations are measured in the
face of high plasma HIV RNA concentrations [60]
.� This appears to be due to diminished fitness of the virus,
resulting from pol gene mutations
selected and maintained by the pressure of HAART.� Circulating T cells exhibited near normal, despite measurable
viremia.� Removal of HAART was deleterious
for the individuals they studied, who experienced a decrease in T cell counts
within two to four weeks, which correlated with the re-emergence of circulating
virus with increased susceptibility (decreased resistance) to reverse
transcriptase and protease inhibitors.�
The destruction rate of CD4+ T cells increased in parallel (Deeks et al
(JID in press)).� They also demonstrated
that HIV with mutations that confer decreased susceptibility to protease
inhibitors also appeared to replicate poorly in thymic organ cultures. These
data suggest that maintenance of a drug resistant genotype and phenotype may be
an acceptable therapeutic goal for selected patients in whom HAART fails to
control HIV replication.� This is likely
to be a fruitful area of investigation in the study of perinatally infected
adolescents, who often have already been treated with a series of therapeutic
regimens that failed to suppress HIV viremia.
Quantitation
of CTL responses
��������������� The rate of progression of HIV related disease in
untreated individuals is clearly related to the level of plasma viremia that
becomes established after acute infection.�
By analogy to homeostatic physiologic mechanisms, this has been referred
to as the viral� �set-point�, which
appears to be linked to the magnitude of CD8 responses to HIV antigens [61] .� Other
studies involving adults have shown that slow progression of disease is
correlated with the presence of potent CTL and CD4+ helper responses [15, 62-64] .�
Until recently, the
mapping of CTL responses has been extremely labor intensive and technically
challenging.� Whole peripheral blood
mononuclear cells (PBMC) from infected persons were randomly cloned at limiting
dilution, followed by screening for HIV-specific cytolytic activity by chromium
release using recombinant vaccinia.�
Alternatively, whole PBMC were specifically stimulated with recombinant
vaccinia-infected autologous B cells (to enrich for HIV-1-specific CTL), then
purified by limiting dilution cloning.�
More recently, the ELISpot assay has been increasingly used to map the
breadth and magnitude of HIV-1 specific cellular immune responses [65, 66] .� Fine-mapping
of CTL responses is possible, due to the great sensitivity of this technique,
which can identify cytokine release from individual cells.� Because synthetic peptides slightly larger
than the optimal epitope can be bound to class I MHC molecules when added
exogenously, mapping of CTL responses within bulk uncloned PBMC can be
accomplished using overlapping synthetic peptides covering the protein of
interest.
This
approach has been recently applied to the study of CTL responses in acute and
chronic HIV.� Goulder et al. used 290
overlapping peptides representing Gag, nef, RT, gp41, gp120, Tat and Rev , and
130 additional peptide represent previously known epitopes to map CTL responses
in 11 subjects with early infection [66]
.� In two
subjects, response to a common HLA-A*0201 restricted epitope were not initially
detected, showing that these responses evolve, and revealing the limitations of
targeting mapping based on HLA type.�
These and other recent studies demonstrate that 1) cellular immune
responses can evolve in a rapid fashion, 2) cannot be dependably predicted on
the basis of MHC restriction [67] , and 3) can be thoroughly examined and quantified
without necessarily knowing the amino acid sequences of autologous virus
sequences.���
Summary.
Detailed
studies of the interplay between HIV infection, thymic output (thymopoiesis),
and antiretroviral therapy, have been critical to recent advances in the
understanding of the pathogenesis of HIV infection and AIDS.� The urgency to better understand the effects
of HIV on thymopoiesis is evident when we consider that HAART has made survival
into adulthood quite likely for the majority of infected infants and children
in the United States and other countries where intensive antiretroviral therapy
is available and accessible. Comprehensive studies of thymopoiesis, and characteristics
of HIV that impair it may guide our efforts to preserve and improved
immunological functions in HIV infected adolescents.� PCR methods to quantify recent thymic emigrants, and in vivo labeling methods to track the
fate of these cells, have made it possible to more comprehensively examine the
mechanistic underpinnings of T cell depletion in pediatric HIV infection.� For the first time, methods now exist to
quantify with some accuracy the production, function, and clearance of T cells.
We do not know what combination of viral, host, and treatment factors has allowed some infants to survive into adolescence after perinatal HIV infection, and can presently only speculate what the future holds in store for them. Taking scientific advantage of the large number of perinatally infected adolescents and young adults in the Los Angeles area, we are proposing studies to examine the balance between the pathogenic properties of HIV, the suppressive and selective power of antiretroviral therapy, and the regenerative capacity of the immune system that exists in these individuals.�
1.������������ Bamji M, Thea DM, Weedon J,
Krasinski K, Matheson PB, Thomas P, Lambert G, Abrams EJ, Steketee R, Heagarty
M. Prospective study of human immunodeficiency virus 1-related disease among
512 infants born to infected women in New York City. The New York City
Perinatal HIV Transmission Collaborative Study Group. Pediatr Infect Dis J
1996;15:891-8.
2.������������ Barnhart HX, Caldwell MB, Thomas P,
Mascola L, Ortiz I, Hsu HW, Schulte J, Parrott R, Maldonado Y, Byers R. Natural
history of human immunodeficiency virus disease in perinatally infected
children: an analysis from the Pediatric Spectrum of Disease Project.
Pediatrics 1996;97:710-6.
3.������������ Galli L, de Martino M, Tovo PA,
Gabiano C, Zappa M. Predictive value of the HIV paediatric classification
system for the long-term course of perinatally infected children. Int J
Epidemiol 2000;29:573-8.
4.������������ CDC. HIV/AIDS Surveillance Report.
End. Vol.12, No.2 ed, 2000
5.������������ Poli G. Cytokines and Chemokines in
HIV Infection. Retroviral Immunology: Immune Response and Restoration Edited
by: Giuseppe Pantaleo and Bruce D Walker 2001; 53-78.
6.������������ Samson M, Libert F, Doranz BJ,
Rucker J, Liesnard C, Farber CM, Saragosti S, Lapoumeroulie C, Cognaux J, Forceille
C, Muyldermans G, Verhofstede C, Burtonboy G, Georges M, Imai T, Rana S, Yi Y,
Smyth RJ, Collman RG, Doms RW, Vassart G, Parmentier M. Resistance to HIV-1
infection in caucasian individuals bearing mutant alleles of the CCR-5
chemokine receptor gene. Nature 1996;382:722-5.
7.������������ Rousseau C, Abrams E, Lee M, Urbano
R, King MC. Long terminal repeat and nef gene variants of human
immunodeficiency virus type 1 in perinatally infected long-term survivors and
rapid progressors. AIDS Res Hum Retroviruses 1997;13:1611-23.
8.������������ Geffin R, Wolf D, Muller R, Hill
MD, Stellwag E, Freitag M, Sass G, Scott GB, Baur AS. Functional and structural
defects in HIV type 1 nef genes derived from pediatric long-term survivors.
AIDS Res Hum Retroviruses 2000;16:1855-68.
9.������������ Nielsen K, McSherry G, Petru A,
Frederick T, Wara D, Bryson Y, Martin N, Hutto C, Ammann AJ, Grubman S, Oleske
J, Scott GB. A descriptive survey of pediatric human immunodeficiency
virus-infected long-term survivors. Pediatrics 1997;99:E4.
10.���������� Ho DD, Neumann AU, Perelson AS, Chen
W, Leonard JM, Markowitz M. Rapid turnover of plasma virions and CD4
lymphocytes in HIV-1 infection. Nature 1995;373:123-6.
11.���������� Wei X, Ghosh SK, Taylor ME, Johnson
VA, Emini EA, Deutsch P, Lifson JD, Bonhoeffer S, Nowak MA, Hahn BH, et al.
Viral dynamics in human immunodeficiency virus type 1 infection. Nature
1995;373:117-22.
12.���������� Autran B, Carcelain G, Li TS, Blanc
C, Mathez D, Tubiana R, Katlama C, Debre P, Leibowitch J. Positive effects of
combined antiretroviral therapy on CD4+ T cell homeostasis and function in
advanced HIV disease. Science 1997;277:112-6.
13.���������� Autran B, Carcelaint G, Li TS,
Gorochov G, Blanc C, Renaud M, Durali M, Mathez D, Calvez V, Leibowitch J,
Katlama C, Debre P. Restoration of the immune system with anti-retroviral
therapy. Immunol Lett 1999;66:207-11.
14.���������� Pakker NG, Notermans DW, de Boer RJ,
Roos MT, de Wolf F, Hill A, Leonard JM, Danner SA, Miedema F, Schellekens PT.
Biphasic kinetics of peripheral blood T cells after triple combination therapy
in HIV-1 infection: a composite of redistribution and proliferation. Nat Med
1998;4:208-14.
15.���������� Pantaleo G, M.D. ; Walker, Bruce D.,
M.D. Retroviral Immunology: Immune Response and Restoration Totowa, New Jersey:
Humana Press, 2001 (Georgiev VS, ed. National Institute of Allergy and
Infectious Diseases. National Institutes of Health)
16.���������� Bohler T, Walcher J, Holzl-Wenig G,
Geiss M, Buchholz B, Linde R, Debatin KM. Early effects of antiretroviral
combination therapy on activation, apoptosis and regeneration of T cells in HIV-1-infected
children and adolescents. Aids 1999;13:779-89.
17.���������� Cohen Stuart JW, Slieker WA, Rijkers
GT, Noest A, Boucher CA, Suur MH, de Boer R, Geelen SP, Scherpbier HJ, Hartwig
NG, Hooijkaas H, Roos MT, de Graeff-Meeder B, de Groot R. Early recovery of CD4+
T lymphocytes in children on highly active antiretroviral therapy. Dutch study
group for children with HIV infections. Aids 1998;12:2155-9.
18.���������� Berkelhamer S, Borock E, Elsen C,
Englund J, Johnson D. Effect of highly active antiretroviral therapy on the
serological response to additional measles vaccinations in human
immunodeficiency virus-infected children. Clin Infect Dis 2001;32:1090-4.
19.���������� Essajee SM, Kim M, Gonzalez C, Rigaud
M, Kaul A, Chandwani S, Hoover W, Lawrence R, Spiegel H, Pollack H, Krasinski
K, Borkowsky W. Immunologic and virologic responses to HAART in severely
immunocompromised HIV-1-infected children. Aids 1999;13:2523-32.
20.���������� Chougnet C, Jankelevich S, Fowke K,
Liewehr D, Steinberg SM, Mueller BU, Pizzo PA, Yarchoan R, Shearer GM. Long-term
protease inhibitor-containing therapy results in limited improvement in T cell
function but not restoration of interleukin-12 production in pediatric patients
with aids. J Infect Dis 2001;184:201-5.
21.���������� Komanduri KV, Viswanathan MN, Wieder
ED, Schmidt DK, Bredt BM, Jacobson MA, McCune JM. Restoration of
cytomegalovirus-specific CD4+ T-lymphocyte responses after ganciclovir and
highly active antiretroviral therapy in individuals infected with HIV-1. Nat
Med 1998;4:953-6.
22.���������� Furrer H, Egger M, Opravil M,
Bernasconi E, Hirschel B, Battegay M, Telenti A, Vernazza PL, Rickenbach M,
Flepp M, Malinverni R. Discontinuation of primary prophylaxis against
Pneumocystis carinii pneumonia in HIV-1-infected adults treated with
combination antiretroviral therapy. Swiss HIV Cohort Study. N Engl J Med
1999;340:1301-6.
23.���������� Connors M, Kovacs JA, Krevat S,
Gea-Banacloche JC, Sneller MC, Flanigan M, Metcalf JA, Walker RE, Falloon J,
Baseler M, Feuerstein I, Masur H, Lane HC. HIV infection induces changes in
CD4+ T-cell phenotype and depletions within the CD4+ T-cell repertoire that are
not immediately restored by antiviral or immune-based therapies. Nat Med
1997;3:533-40.
24.���������� Gea-Banacloche JC, Martino L, Mican
JM, Hallahan CW, Baseler M, Stevens R, Lambert L, Polis M, Lane HC, Connors M.
Longitudinal changes in CD4+ T cell antigen receptor diversity and naive/memory
cell phenotype during 9 to 26 months of antiretroviral therapy of HIV-infected
patients. AIDS Res Hum Retroviruses 2000;16:1877-86.
25.���������� Gorochov G, Neumann AU, Kereveur A,
Parizot C, Li T, Katlama C, Karmochkine M, Raguin G, Autran B, Debre P.
Perturbation of CD4+ and CD8+ T-cell repertoires during progression to AIDS and
regulation of the CD4+ repertoire during antiviral therapy. Nat Med
1998;4:215-21.
26.���������� McIntosh K, Shevitz A, Zaknun D,
Kornegay J, Chatis P, Karthas N, Burchett SK. Age- and time-related changes in
extracellular viral load in children vertically infected by human
immunodeficiency virus. Pediatr Infect Dis J 1996;15:1087-91.
27.���������� Dickover RE, Dillon M, Leung KM,
Krogstad P, Plaeger S, Kwok S, Christopherson C, Deveikis A, Keller M, Stiehm
ER, Bryson YJ. Early prognostic indicators in primary perinatal human
immunodeficiency virus type 1 infection: importance of viral RNA and the timing
of transmission on long-term outcome. J Infect Dis 1998;178:375-87.
28.���������� Kourtis AP, Ibegbu C, Nahmias AJ, Lee
FK, Clark WS, Sawyer MK, Nesheim S. Early progression of disease in
HIV-infected infants with thymus dysfunction. N Engl J Med 1996;335:1431-6.
29.���������� Nahmias AJ, Clark WS, Kourtis AP, Lee
FK, Cotsonis G, Ibegbu C, Thea D, Palumbo P, Vink P, Simonds RJ, Nesheim SR.
Thymic dysfunction and time of infection predict mortality in human
immunodeficiency virus-infected infants. CDC Perinatal AIDS Collaborative Transmission
Study Group. J Infect Dis 1998;178:680-5.
30.���������� Haynes BF, Markert ML, Sempowski GD,
Patel DD, Hale LP. The role of the thymus in immune reconstitution in aging,
bone marrow transplantation, and HIV-1 infection. Annu Rev Immunol
2000;18:529-60
31.���������� Withers-Ward ES, Amado RG, Koka PS,
Jamieson BD, Kaplan AH, Chen IS, Zack JA. Transient renewal of thymopoiesis in
HIV-infected human thymic implants following antiviral therapy. Nat Med
1997;3:1102-9.
32.���������� Viard JP, Mocroft A, Chiesi A, Kirk
O, Roge B, Panos G, Vetter N, Bruun JN, Johnson M, Lundgren JD. Influence of
age on CD4 cell recovery in human immunodeficiency virus- infected patients
receiving highly active antiretroviral therapy: evidence from the EuroSIDA
study. J Infect Dis 2001;183:1290-4.
33.���������� Franco JM, Leon-Leal JA, Leal M,
Cano-Rodriguez A, Pineda JA, Macias J, Rubio A, Rey C, Sanchez B, Lissen E.
CD4+ and CD8+ T lymphocyte regeneration after anti-retroviral therapy in
HIV-1-infected children and adult patients. Clin Exp Immunol 2000;119:493-8.
34.���������� Steinmann GG. Changes in the human
thymus during aging. Curr Top Pathol 1986;75:43-88
35.���������� Flores KG, Li J, Sempowski GD, Haynes
BF, Hale LP. Analysis of the human thymic perivascular space during aging. J
Clin Invest 1999;104:1031-9.
36.���������� Jamieson BD, Douek DC, Killian S,
Hultin LE, Scripture-Adams DD, Giorgi JV, Marelli D, Koup RA, Zack JA.
Generation of functional thymocytes in the human adult. Immunity
1999;10:569-75.
37.���������� Douek DC, McFarland RD, Keiser PH,
Gage EA, Massey JM, Haynes BF, Polis MA, Haase AT, Feinberg MB, Sullivan JL,
Jamieson BD, Zack JA, Picker LJ, Koup RA. Changes in thymic function with age
and during the treatment of HIV infection. Nature 1998;396:690-5.
38.���������� Brooks DG, Kitchen SG, Kitchen CM,
Scripture-Adams DD, Zack JA. Generation of HIV latency during thymopoiesis. Nat
Med 2001;7:459-64.
39.���������� Pierson T, Hoffman TL, Blankson J,
Finzi D, Chadwick K, Margolick JB, Buck C, Siliciano JD, Doms RW, Siliciano RF.
Characterization of chemokine receptor utilization of viruses in the latent
reservoir for human immunodeficiency virus type 1. J Virol 2000;74:7824-33.
40.���������� Mahoney EM, Donfield SM, Howard C,
Kaufman F, Gertner JM. HIV-associated immune dysfunction and delayed pubertal
development in a cohort of young hemophiliacs. Hemophilia Growth and
Development Study. J Acquir Immune Defic Syndr 1999;21:333-7.
41.���������� Linton P, Thoman ML. T cell
senescence. Front Biosci 2001;6:D248-61.
42.���������� Sfikakis PP, Kostomitsopoulos N,
Kittas C, Stathopoulos J, Karayannacos P, Dellia-Sfikakis A, Mitropoulos D.
Tamoxifen exerts testosterone-dependent and independent effects on thymic
involution. Int J Immunopharmacol 1998;20:305-12.
43.���������� Moore AV, Korobkin M, Olanow W,
Heaston DK, Ram PC, Dunnick NR, Silverman PM. Age-related changes in the thymus
gland: CT-pathologic correlation. AJR Am J Roentgenol 1983;141:241-6.
44.���������� McCune JM, Loftus R, Schmidt DK,
Carroll P, Webster D, Swor-Yim LB, Francis IR, Gross BH, Grant RM. High
prevalence of thymic tissue in adults with human immunodeficiency virus-1
infection. J Clin Invest 1998;101:2301-8.
45.���������� McCune JM, Hanley MB, Cesar D,
Halvorsen R, Hoh R, Schmidt D, Wieder E, Deeks S, Siler S, Neese R, Hellerstein
M. Factors influencing T-cell turnover in HIV-1-seropositive patients. J Clin
Invest 2000;105:R1-8.
46.���������� Vigano A, Vella S, Saresella M,
Vanzulli A, Bricalli D, Di Fabio S, Ferrante P, Andreotti M, Pirillo M, Dally
LG, Clerici M, Principi N. Early immune reconstitution after potent
antiretroviral therapy in HIV- infected children correlates with the increase
in thymus volume. Aids 2000;14:251-61.
47.���������� Kong F, Chen CH, Cooper MD. Thymic
function can be accurately monitored by the level of recent T cell emigrants in
the circulation. Immunity 1998;8:97-104.
48.���������� Kong FK, Chen CL, Six A, Hockett RD,
Cooper MD. T cell receptor gene deletion circles identify recent thymic
emigrants in the peripheral T cell pool. Proc Natl Acad Sci U S A
1999;96:1536-40.
49.���������� Zhang L, Lewin SR, Markowitz M, Lin
HH, Skulsky E, Karanicolas R, He Y, Jin X, Tuttleton S, Vesanen M, Spiegel H,
Kost R, van Lunzen J, Stellbrink HJ, Wolinsky S, Borkowsky W, Palumbo P,
Kostrikis LG, Ho DD. Measuring recent thymic emigrants in blood of normal and
HIV-1-infected individuals before and after effective therapy. J Exp Med
1999;190:725-32.
50.���������� Poulin JF, Viswanathan MN, Harris JM,
Komanduri KV, Wieder E, Ringuette N, Jenkins M, McCune JM, Sekaly RP. Direct
evidence for thymic function in adult humans. J Exp Med 1999;190:479-86.
51.���������� De Villartay JP, Hockett RD, Coran D,
Korsmeyer SJ, Cohen DI. Deletion of the human T-cell receptor delta-gene by a
site-specific recombination. Nature 1988;335:170-4.
52.���������� Fujimoto S, Yamagishi H. Isolation of
an excision product of T-cell receptor alpha-chain gene rearrangements. Nature
1987;327:242-3.
53.���������� Livak F, Schatz DG. T-cell receptor
alpha locus V(D)J recombination by-products are abundant in thymocytes and
mature T cells. Mol Cell Biol 1996;16:609-18.
54.���������� Chavan S, Bennuri B, Kharbanda M,
Chandrasekaran A, Bakshi S, Pahwa S. Evaluation of T cell receptor gene
rearrangement excision circles after antiretroviral therapy in children
infected with human immunodeficiency virus. J Infect Dis 2001;183:1445-54.
55.���������� Hazenberg MD, Otto SA, Cohen Stuart
JW, Verschuren MC, Borleffs JC, Boucher CA, Coutinho RA, Lange JM, Rinke de Wit
TF, Tsegaye A, van Dongen JJ, Hamann D, de Boer RJ, Miedema F. Increased cell
division but not thymic dysfunction rapidly affects the T-cell receptor
excision circle content of the naive T cell population in HIV-1 infection. Nat
Med 2000;6:1036-42.
56.���������� Grossman Z, Paul WE. The impact of
HIV on naive T-cell homeostasis. Nat Med 2000;6:976-7.
57.���������� Tanchot C, Rocha B. The organization
of mature T-cell pools. Immunol Today 1998;19:575-9.
58.���������� Freitas AA, Rocha B. Peripheral T
cell survival. Curr Opin Immunol 1999;11:152-6.
59.���������� Hellerstein M, Hanley MB, Cesar D,
Siler S, Papageorgopoulos C, Wieder E, Schmidt D, Hoh R, Neese R, Macallan D,
Deeks S, McCune JM. Directly measured kinetics of circulating T lymphocytes in
normal and HIV-1-infected humans. Nat Med 1999;5:83-9.
60.���������� Deeks SG, Wrin T, Liegler T, Hoh R,
Hayden M, Barbour JD, Hellmann NS, Petropoulos CJ, McCune JM, Hellerstein MK,
Grant RM. Virologic and immunologic consequences of discontinuing combination
antiretroviral-drug therapy in HIV-infected patients with detectable viremia. N
Engl J Med 2001;344:472-80.
61.���������� Ogg GS, Jin X, Bonhoeffer S, Dunbar
PR, Nowak MA, Monard S, Segal JP, Cao Y, Rowland-Jones SL, Cerundolo V, Hurley
A, Markowitz M, Ho DD, Nixon DF, McMichael AJ. Quantitation of HIV-1-specific
cytotoxic T lymphocytes and plasma load of viral RNA. Science 1998;279:2103-6.
62.���������� Klein MR, van Baalen CA, Holwerda AM,
Kerkhof Garde SR, Bende RJ, Keet IP, Eeftinck-Schattenkerk JK, Osterhaus AD,
Schuitemaker H, Miedema F. Kinetics of Gag-specific cytotoxic T lymphocyte
responses during the clinical course of HIV-1 infection: a longitudinal
analysis of rapid progressors and long-term asymptomatics. J Exp Med
1995;181:1365-72.
63.���������� Rinaldo CR, Jr., Beltz LA, Huang XL,
Gupta P, Fan Z, Torpey DJ, 3rd. Anti-HIV type 1 cytotoxic T lymphocyte effector
activity and disease progression in the first 8 years of HIV type 1 infection
of homosexual men. AIDS Res Hum Retroviruses 1995;11:481-9.
64.���������� Harrer T, Harrer E, Kalams SA,
Barbosa P, Trocha A, Johnson RP, Elbeik T, Feinberg MB, Buchbinder SP, Walker
BD. Cytotoxic T lymphocytes in asymptomatic long-term nonprogressing HIV-1
infection. Breadth and specificity of the response and relation to in vivo
viral quasispecies in a person with prolonged infection and low viral load. J
Immunol 1996;156:2616-23.
65.���������� Altfeld M, Rosenberg ES, Shankarappa
R, Mukherjee JS, Hecht FM, Eldridge RL, Addo MM, Poon SH, Phillips MN, Robbins
GK, Sax PE, Boswell S, Kahn JO, Brander C, Goulder PJ, Levy JA, Mullins JI,
Walker BD. Cellular immune responses and viral diversity in individuals treated
during acute and early HIV-1 infection. J Exp Med 2001;193:169-80.
66.���������� Goulder PJ, Altfeld MA, Rosenberg ES,
Nguyen T, Tang Y, Eldridge RL, Addo MM, He S, Mukherjee JS, Phillips MN, Bunce
M, Kalams SA, Sekaly RP, Walker BD, Brander C. Substantial differences in
specificity of HIV-specific cytotoxic T cells in acute and chronic HIV
infection. J Exp Med 2001;193:181-94.
67.���������� Betts MR, Casazza JP, Patterson BA,
Waldrop S, Trigona W, Fu TM, Kern F, Picker LJ, Koup RA. Putative immunodominant
human immunodeficiency virus-specific CD8(+) T- cell responses cannot be
predicted by major histocompatibility complex class I haplotype. J Virol
2000;74:9144-51.
68.���������� Misrahi M, Teglas JP, N'Go N, Burgard
M, Mayaux MJ, Rouzioux C, Delfraissy JF, Blanche S. CCR5 chemokine receptor
variant in HIV-1 mother-to-child transmission and disease progression in
children. French Pediatric HIV Infection Study Group. Jama 1998;279:277-80.
69.���������� Douek DC, Vescio RA, Betts MR,
Brenchley JM, Hill BJ, Zhang L, Berenson JR, Collins RH, Koup RA. Assessment of
thymic output in adults after haematopoietic stem-cell transplantation and
prediction of T-cell reconstitution. Lancet 2000;355:1875-81.
70.���������� Rogers AS, Futterman DK, Moscicki AB,
Wilson CM, Ellenberg J, Vermund SH. The REACH Project of the Adolescent
Medicine HIV/AIDS Research Network: design, methods, and selected
characteristics of participants. J Adolesc Health 1998;22:300-11.
71.���������� Vigano A, Vella S, Principi N,
Bricalli D, Sala N, Salvaggio A, Saresella M, Vanzulli A, Clerici M. Thymus
volume correlates with the progression of vertical HIV infection. Aids
1999;13:F29-34.
72.���������� Baron RL, Lee JK, Sagel SS, Peterson
RR. Computed tomography of the normal thymus. Radiology 1982;142:121-5.
73.���������� Brown MS, McNitt-Gray MF, Mankovich
NJ, Goldin JG, Hiller J, Wilson LS, Aberle DR. Method for segmenting chest CT
image data using an anatomical model: preliminary results. IEEE Trans Med
Imaging 1997;16:828-39.
74.���������� Brown MS, McNitt-Gray MF, Goldin JG,
Greaser LE, Hayward UM, Sayre JW, Arid MK, Aberle DR. Automated measurement of
single and total lung volume from CT. J Comput Assist Tomogr 1999;23:632-40.
75.���������� Brown MS, Feng WC, Hall TR,
McNitt-Gray MF, Churchill BM. Knowledge-based segmentation of pediatric kidneys
in CT for measurement of parenchymal volume. J Comput Assist Tomogr
2001;25:639-48.
76.���������� Gregersen PK, Hingorani R, Monteiro
J. Oligoclonality in the CD8+ T-cell population. Analysis using a multiplex PCR
assay for CDR3 length. Ann N Y Acad Sci 1995;756:19-27.
77.���������� Kou ZC, Puhr JS, Rojas M, McCormack
WT, Goodenow MM, Sleasman JW. T-Cell receptor Vbeta repertoire CDR3 length
diversity differs within CD45RA and CD45RO T-cell subsets in healthy and human
immunodeficiency virus-infected children. Clin Diagn Lab Immunol 2000;7:953-9.
78.���������� Killian S, Monteiro, Joanita,���� Matud, Jose,� Hultin, Lance,� Hausner,
Mary Ann,� Jamieson, Beth D.� Gregersen, Peter K.� Detels,Roger,� Giorgi, Janis V. History of Antigenic Exposure Evident in the
T-Cell Repertoire. Submitted.
79.���������� Walker BD, Korber BT. Immune control
of HIV: the obstacles of HLA and viral diversity. Nat Immunol 2001;2:473-5.
80.���������� Delwart EL, Gordon CJ. Tracking
changes in HIV-1 envelope quasispecies using DNA heteroduplex analysis. Methods
1997;12:348-54.
81.���������� Dickover RE, Garratty EM, Plaeger S,
Bryson YJ. Perinatal transmission of major, minor, and multiple maternal human
immunodeficiency virus type 1 variants in utero and intrapartum. J Virol
2001;75:2194-203.
82.���������� Wiznia A, Stanley K, Krogstad P,
Johnson G, Lee S, McNamara J, Moye J, Jackson JB, Mendez H, Aguayo R, Dieudonne
A, Kovacs A, Bamji M, Abrams E, Rana S, Sever J, Nachman S. Combination
nucleoside analog reverse transcriptase inhibitor(s) plus nevirapine,
nelfinavir, or ritonavir in stable antiretroviral therapy- experienced
HIV-infected children: week 24 results of a randomized controlled trial--PACTG
377. Pediatric AIDS Clinical Trials Group 377 Study Team. AIDS Res Hum
Retroviruses 2000;16:1113-21.
83.���������� Cunningham S, Ank B, Lewis D, Lu W,
Wantman M, Dileanis JA, Jackson JB, Palumbo P, Krogstad P, Eshleman SH.
Performance of the applied biosystems ViroSeq human immunodeficiency virus type
1 (HIV-1) genotyping system for sequence-based analysis of HIV-1 in pediatric
plasma samples. J Clin Microbiol 2001;39:1254-7.
84.���������� Eshleman SH, Krogstad P, Jackson JB,
Wang YG, Lee S, Wei LJ, Cunningham S, Wantman M, Wiznia A, Johnson G, Nachman
S, Palumbo P. Analysis of human immunodeficiency virus type 1 drug resistance
in children receiving nucleoside analogue reverse-transcriptase inhibitors plus
nevirapine, nelfinavir, or ritonavir (Pediatric AIDS Clinical Trials Group
377). J Infect Dis 2001;183:1732-8.
85.���������� Bryson YJ, Luzuriaga K, Sullivan JL,
Wara DW. Proposed definitions for in utero versus intrapartum transmission of
HIV-1. N Engl J Med 1992;327:1246-7.
86.���������� Pedroza-Martins L, Gurney KB, Torbett
BE, Uittenbogaart CH. Differential tropism and replication kinetics of human
immunodeficiency virus type 1 isolates in thymocytes: coreceptor expression
allows viral entry, but productive infection of distinct subsets is determined
at the postentry level. J Virol 1998;72:9441-52.
87.���������� www.hivatis.org
88.���������� Napolitano
LA, Grant RM, Deeks SG, Schmidt D, De Rosa SC, Herzenberg LA, Herndier BG,
Andersson J, McCune JM. Increased
production of IL-7 accompanies HIV-1-mediated T-cell depletion: implications
for T-cell homeostasis. Nat Med 2001;7:73-9.
89.���������� Ferbas J, Kaplan AH, Hausner MA,
Hultin LE, Matud JL, Liu Z, Panicali DL, Nerng-Ho H, Detels R, Giorgi JV. Virus
burden in long-term survivors of human immunodeficiency virus (HIV) infection
is a determinant of anti-HIV CD8+ lymphocyte activity. J Infect Dis
1995;172:329-39.
90.���������� Equils O, Garratty E, Wei LS, Plaeger
S, Tapia M, Deville J, Krogstad P, Sim MS, Nielsen K, Bryson YJ. Recovery of
replication-competent virus from CD4 T cell reservoirs and change in coreceptor
use in human immunodeficiency virus type 1- infected children responding to
highly active antiretroviral therapy. J Infect Dis 2000;182:751-7.
91.���������� Chun TW, Finzi D, Margolick J,
Chadwick K, Schwartz D, Siliciano RF. In vivo fate of HIV-1-infected T cells:
quantitative analysis of the transition to stable latency. Nat Med
1995;1:1284-90.
92.���������� Zack JA, Haislip AM, Krogstad P, Chen
IS. Incompletely reverse-transcribed human immunodeficiency virus type 1
genomes in quiescent cells can function as intermediates in the retroviral life
cycle. J Virol 1992;66:1717-25.
93.���������� Myers LE, McQuay LJ, Hollinger FB.
Dilution assay statistics. J Clin Microbiol 1994;32:732-9.
94.���������� Lee S, Tiffany HL, King L, Murphy PM,
Golding H, Zaitseva MB. CCR8 on human thymocytes functions as a human
immunodeficiency virus type 1 coreceptor. J Virol 2000;74:6946-52.
95.���������� Delwart EL, Pan H, Sheppard HW,
Wolpert D, Neumann AU, Korber B, Mullins JI. Slower evolution of human immunodeficiency
virus type 1 quasispecies during progression to AIDS. J Virol 1997;71:7498-508.
96.���������� Aldrovandi GM, Feuer G, Gao L,
Jamieson B, Kristeva M, Chen IS, Zack JA. The SCID-hu mouse as a model for
HIV-1 infection. Nature 1993;363:732-6.
97.���������� Stanley SK, McCune JM, Kaneshima H,
Justement JS, Sullivan M, Boone E, Baseler M, Adelsberger J, Bonyhadi M,
Orenstein J, et al. Human immunodeficiency virus infection of the human thymus
and disruption of the thymic microenvironment in the SCID-hu mouse. J Exp Med 1993;178:1151-63.
98.���������� Bonyhadi ML, Rabin L, Salimi S, Brown
DA, Kosek J, McCune JM, Kaneshima H. HIV induces thymus depletion in vivo.
Nature 1993;363:728-32.
99.���������� Boldt-Houle DM, Jamieson BD,
Aldrovandi GM, Rinaldo CR, Jr., Ehrlich GD, Zack JA. Loss of T cell receptor
Vbeta repertoires in HIV type 1-infected SCID- hu mice. AIDS Res Hum
Retroviruses 1997;13:125-34.
100.�������� Yang OO, Kalams SA, Trocha A, Cao H,
Luster A, Johnson RP, Walker BD. Suppression of human immunodeficiency virus
type 1 replication by CD8+ cells: evidence for HLA class I-restricted
triggering of cytolytic and noncytolytic mechanisms. J Virol 1997;71:3120-8.
101.�������� Rosenberg
ES, Altfeld M, Poon SH, Phillips MN, Wilkes BM, Eldridge RL, Robbins GK,
D'Aquila RT, Goulder PJ, Walker BD. Immune control of HIV-1 after early
treatment of acute infection. Nature 2000;407:523-6.
CHARACTERISTICS OF THE SUBJECT POPULATION (4-6) |
4. ��� Number of Subjects:� What is the anticipated number of
subjects to be enrolled at UCLA and, in the case of multi-center research, the
total number of subjects for the entire project?
A total of 60-90 adolescents and young adults will be
enrolled.��� There will be 20-30
subjects from each group ( perinatally infected = PI-A; seronegative = SN-A;
and adult behavior infected = AB-A).
5. ��� Inclusion/Exclusion Criteria: �
������� a)���� What are the criteria for inclusion and
exclusion?�
������� b)��� How will eligibility be determined, and by
whom?�
c)���� Are any
inclusion or exclusion criteria based on age, gender, pregnancy or childbearing
potential, or racial/ethnic origin?� If
so, explain and justify.
������� Note: Equitable inclusion of both men
and women of all ages, and individuals from diverse racial/ethnic backgrounds,
is important to assure that they receive an equal share of the benefits of
research and that they do not bear a disproportionate share of its
burdens.� Participation of adult
subjects of both genders and diverse racial/ethnic backgrounds should not be
restricted without medical or scientific justification.
a.
The study is open to subjects between the ages of 13
and 21 at the time of enrollment, irrespective of race and gender.� Given the potential 30 month study period,
all subjects will complete the study at the age of 24 or younger.��� 20-30 HIV positive subjects, perinatally
infected subjects will be recruited, along with 20-30� subjects seronegative for HIV and 20-30 HIV positive subjects who
acquired HIV through adult behavior.
b.
Subject eligibility will be determined by the
principal investigator based on the above criteria
c.
Subjects 12 and younger will be excluded since this
study will focus on HIV pathogenesis in pubertal and post pubertal children.
Subjects greater than 21 at the time of screening will be excluded for the same
reason.
6. ��� Vulnerable Subjects:� Will any vulnerable subjects be included? If
so, identify the subject groups and justify their involvement.
Examples of vulnerable subjects: children, elderly, pregnant women,
fetuses, cognitively impaired individuals, persons with severe psychological
disorders, terminally ill patients, emergency patients, institutional
residents, prisoners, parolees, non-English speaking subjects, and UCLA
students/staff.
Adolescents
and young adults will be the sole subjects, since the study is designed to
focus on HIV pathogenesis in pubertal and post pubertal children.
SUBJECT IDENTIFICATION AND RECRUITMENT (7) |
7. ��� Method of Subject Identification and
Recruitment: �What method(s) will be
used to identify and recruit prospective subjects? Attach a copy of any
planned advertisements/notices and letters to potential subjects.
������� Note: The identification and recruitment of
subjects must be ethically and legally acceptable and free of coercion.� Procedures used to recruit subjects should
be designed to reach diverse populations.�
Vulnerable subjects, such as persons in nursing homes or institutions,
should not be recruited merely for the sake of convenience.
Subjects will be recruited through the use of flyers
(see attached).� In addition, the
investigator will let his colleagues within UCLA know about the existence of
the study and will give them flyers such that interested subjects may contact
the investigator to learn more about the study.
METHODS AND PROCEDURES
(8-11) |
8. ��� Methods and Procedures Applied to Human
Subjects:� Describe the study design
and all procedures (sequentially) to which human participants will be
subjected. Identify all procedures that are considered experimental and/or
procedures performed exclusively for research purposes.
������� Note: A clinical research protocol may
involve interventions that are strictly experimental or it may involve some
aspect of research (e.g., randomization among standard treatments for
collection and analysis of routine clinical data for research purposes). It is
important for this section to distinguish between interventions that are
experimental and/or carried out for research purposes versus those procedures
that are considered standard therapy. In addition, routine procedures performed
solely for research purposes (e.g., additional diagnostic/follow-up tests)
should be identified.
Month 0
After consenting/assenting
to participate in the study, subjects will have a physical exam and will have
blood collected.� They will also have a
CT scan.
Month 6
Subjects will have a
physical exam and will have blood collected.�
They may also be asked if they want to participate in the substudy (see
below).
Months 12 and 18
Control (non HIV subjects)
will have a physical exam and will have blood collected.� The study ends for control subjects at month
18
HIV infected subjects will
have a physical exam and will have blood collected.� Depending on the results of their month 12 blood work, they may
have a CT scan at the month 18 visit.
Months 24 and 30
Only HIV infected subjects
who have changed their anti-HIV medication regimen will have visits at month 24
and month 30.� At these visits, they
will have blood collected, a physical exam, and a may have CT scan.
At
month 6, subjects may be asked if they would like to pariticipate in the
substudy.� If they are interested, they
will consented/assented using a separate consent/assent.� Subjects would be admitted to the GCRC.� They will be infused with a deuterium
labeled glucose over a 24 hour period.�
A fingerstick for glucose monitoring will be done at 12 and 24
hours.� The subject will be asked to
return to the clinic between 4 and 7 days, and then again between 8 and 14 days
for blood collection (up to 50 ml at each visit, but dependent on body
weight).� It is uncertain if this
glucose infusion will allow adequate labeling.�
If it does not, subsequent subjects would be switched to receive 70% D2O
over 24 hours.� They will then be sent
home with pre-measured aliquots of D2O to drink 2-3 times per week,
and they will be asked to return to the clinic for blood samples at days 14 and
days 28 (up to 50 ml at each visit, but dependent on body weight)
9. ��� For Research Involving Survey,
Questionnaires, etc.: Describe the setting and mode of administering the
instrument (e.g., by telephone, one-on-one, or group) and the provisions for
maintaining privacy and confidentiality.�
Include the duration, intervals of administration, and overall length of
participation.�
������� Note:�
If the protocol for the interviews or the questionnaires are not yet
designed, provide a sample of the questions or describe the subject matter to
be covered.� (If the instrument has been
prepared even in draft form, submit a copy.)�
The final survey instruments or questionnaires must be reviewed and
approved by the HSPC before they may be used.
Not applicable
10. � FDA Approval:� Are there any investigational drugs or biological agents used in
this study?� If yes, please complete
Section IV.� Are there any investigational
devices used in this study?� If yes,
please complete Section V.� If the study
does not involve any investigational drugs or devices, this should be stated.
Not applicable
11. � Data Collection, Storage and
Confidentiality:�
a)���� How will
data be collected and recorded?� Will it
be associated with personal identifiers or coded to
������� protect
personal privacy?
������� b)��� Where will the data be stored during the
study and how will it be secured?
c)���� Who will
have access to the data and/or to the codes? If data with subject identifiers
will be released, specify the person(s) or agency to whom this information will
be released.
������� d)��� What will happen to the data when the
research has been completed?
������� Note: The principal investigator is
responsible for taking all necessary steps to maintain confidentiality� of data. This includes coding data and
choosing an appropriate and secure data storage mechanism that will prevent
unauthorized access to the data.� Where
appropriate, the principal investigator should seek a certificate of
confidentiality from the federal government.
a.
Data will be collected
and recorded on forms designed by the study team.� These forms will be coded to protect personal privacy.
b.
All data will be kept
confidential and will be stored in locked storage in the office of the
investigator.
c.
Only the investigator
and his study team will have access to the data.
d.
The data will be stored
by the principal investigator.
RISK/BENEFIT ASSESSMENT (12-17) |
12. � Potential Risks and Discomforts:� What are the potential risks/discomforts
associated with each intervention or research procedure?� If data are available, estimate (a) the
probability that a given harm may occur, (b) its severity, and (c) its
potential reversibility.
������� Note: A risk/discomfort is a potential harm
associated with the research that a reasonable person would consider important
in deciding whether to participate in the research.� Risks can be generally categorized as physical, psychological,
sociological, economic and legal.
Subjects
will be exposed to a small amount of radiation from the CT scans.� The number of CT scans will be kept to a
minimum, with repeat scans done only on HIV infected subjects who have changed
their HIV medications or whose blood work indicates changes in their immune
status.
Pain,
bruising, rarely infection or fainting may occur
Additional
risks of Substudy:
Pain,
bruising, rarely infection or fainting may occur
The
stay may cause inconvenience to the subject and his or her family.
13. � Risk Classification:� What is the overall risk classification of
the research: minimal, greater than minimal, significant, or unknown?
������� Note: According to HHS/FDA Regulations
minimal risk means "The probability and magnitude of harm or discomfort
anticipated in the research are not greater in and of themselves than those
ordinarily encountered in daily life or during the performance of routine
physical or psychological examinations or tests."� When the risks associated with a new
procedure or product are unknown, they cannot be classified as �minimal.�� Your estimation of risk determines the
Emergency Care and Compensation for research-related Injury clause in the
informed consent form.
Greater than minimal
14. � Minimizing Risks: �What procedure(s) will be utilized to
prevent/minimize any potential risks or discomfort?
������� Note: All potential risks and discomfort
must be minimized to the greatest extent possible by using procedures such as
appropriate monitoring and withdrawal of the subject upon evidence of a
specific adverse event or clinical sign(s). This section should reflect that
all appropriate steps will be taken to protect subjects from harm.
All procedures will be performed by trained medical
personnel.� CT scans after the month 0
visit will be done only on subjects who have altered their HIV medications or
whose blood work shows changes in their immune status.� Blood volumes collected will be based on
subject�s body weight to ensure the guidelines regarding maximum collection are
not exceeded.�
15. � Potential Benefits: �
a)���� What
potential benefits may subjects receive as a result of their participation in
the research?
b)��� What potential
benefits to society may be expected from this research?
������� Note: Societal benefits� generally refer to the advancement of
medical knowledge and/or possible benefit to future patients
a.
Subjects will not
benefit from their participation in this study.
b.
The study results may
give more information on the affects of HIV infection on the immune status and
the thymus, leading to better treatments for HIV infection.
16. � Therapeutic Alternatives:� What therapeutic alternatives are
reasonably available in the non-research and/or research context that may be of
benefit to the potential subjects?
������� Note: This section should include a
reasonably detailed description of the therapeutic alternatives that could be
used to treat the patient should they elect not to participate in the protocol.
Not applicable � the study is not designed to provide
therapeutic intervention.� The
alternative to participation is not to participate.
17. � Risk/Benefit Ratio: What is the
risk/benefit ratio of the research, compared with that of the available
alternatives?�
������� Note: The potential benefits of research
must justify the risks to human subjects.�
Some risks may not be reasonable, no matter how important the potential
benefits.� The risk/benefit ratio of the
research must be at least as favorable for the subjects as that presented by
standard treatments for their condition.�
When comparing the risk/benefit ratio of research with that of available
alternatives, the alternative of doing nothing, or �watchful waiting,� should
be included in the analysis.
The risks of the study are
minimal.� Blood collection and CT scans
are routine procedures that present minimal risk to the subjects.� The pathogenesis of HIV infection and its affect
on the thymus through puberty and immediately post puberty are not well
understood.� This study may give
valuable information regarding this, thus potentially leading to more effective
treatments.� Thus we feel that the
benefits outweigh the risks.
FINANCIAL CONSIDERATIONS (18-20) |
18. � Payment for Participation:� Describe all plans to pay subjects, in cash
or in kind.� If no payment is planned,
that should be stated.� Information
regarding payment consideration should include: Will subjects receive any
financial inducement or payment for participation?� Will they receive services or other benefits instead of
cash?� Will they be reimbursed for
travel and other expenses?� What
conditions must be fulfilled by subjects to receive either full or partial
payment?
������� Note: The FDA encourages a prorated
system of payment whereby subjects who do not finish the protocol are paid in
proportion to the part completed.� The
amount of payment must be justified and not constitute undue inducement of the
subject to participate in the research. If a non-prorated system of payment
will be used, this should be justified in this section.
Control subjects will be paid $20 per visit for the
main study. HIV infected subjects will be paid $10 per visit for the main
study.� This is less than the control
subjects, since the HIV subjects will likely have clinical visits for routine
care at the same frequency as the study visits, and we will attempt to perform
the study visits in conjuction with the clinic visits to maximize convenience
for the subjects.� For the substudy,
subjects will be paid $75 for the overnight visit, $25 for the first visit for
blood collection, and $50 for the second visit for blood collection.
19.�� Financial Obligations of the Subjects:� What financial obligations will subjects
incur as a result of participating in the study?� Will subjects have to pay for any of the treatment(s) they
receive or tests performed in the research?
������� Note: This section should clarify who will
pay for procedures associated with the study as well as financial responsibility
for routine clinical care (e.g., Diagnostic tests, hospitalization,
follow-up).� Insurance and other third
party payers may not cover procedures associated with participation in research
(even if they might have paid for some of the procedures in connection with
standard therapy).� Consequently,
subjects� costs may be increased as a result of additional follow-up
examinations and/or tests required by the research.
There will be no charge to the subjects for
participation in this study.� Subjects
will not be charged for their office visits, physical exams, blood draws, lab
work, CT scans, overnight stay in the GCRC, the labeled glucose or water.
20.�� Emergency Care and Compensation for
Research-Related Injury:� If the
research presents greater than minimal risk, what emergency care is available
in case of research-related injury?� Who
will be responsible for the cost of such care?�
Will subjects be compensated for out-of-pocket expenses or lost wages if
they suffer a research-related injury?
������� Note: Standard language for explaining
this to prospective subjects is provided in the instructions for preparing the
Consent Form. (Forms HS-2 & HS-3).�
Subjects who are injured as a result of research
procedures not done primarily for their own benefit will receive treatment at
no cost.
INFORMED CONSENT (21-26) |
21. � Capacity to Consent:� Will all adult subjects have the
capacity to give informed consent?� If
not, describe the likely range of impairment and explain how, and by whom,
their capacity to consent will be determined.
������� Note: In research involving more than
minimal risk, capacity to consent should be determined by a psychiatrist,
clinical psychologist, or other qualified professional not otherwise involved
in the research.� Individuals who lack
the capacity to consent may participate in research only if consent is given on
their behalf by a legally authorized representative.
Subjects 18 and older will be given the consent
form.� Subjects younger than 18 will be
given the youth assent form and their parents will be given the consent
form.�� All subjects will have the
capacity to consent/assent
22.�� Personnel Inviting Participants:� Who will be inviting subjects to participate
and what will they say?� Identify by
name and training the individual(s) authorized to describe the research to
subjects/representatives and to invite their participation.�
������� Note:�
Only those individuals authorized to solicit consent may sign the
consent form confirming that the prospective subject was provided the necessary
information and that any questions asked were answered.
The investigator will invite all subjects to
participate and will invite the parents/guardians to consent for adolescents
13-17 years old.��� He will describe the
study to the subjects and will reiterate that their participation is completely
voluntary.
23. � Process
of Consent: �How and where will the
consent process take place? How will it be structured to enhance independent
and thoughtful decision-making?� What
steps will be taken to avoid coercion or undue influence?
������� Note:�
Consider:� a) the environment and
location where informed consent will be solicited; b) the timing of the process
(e.g., in relation to hospital admission, surgery, medication, stressful
events); c) the involvement of someone other than the investigators to help
explain the research; and d) opportunity for the� prospective subjects/representatives to discuss participation in
the research with family, friends, or their advisors before signing the consent
form.
The consent will be obtained immediately after the
subject determines that s/he is interested in possible participation in the
study.� The investigator will review the
consent/assent form with the subject and the consent form with the parent/guardian
as applicable.� The subject and
parent/guardian will be given ample opportunities to ask questions and will be
questioned to ensure his/her understanding of the information in the
consent/assent form.� The subject will
be encouraged to take the consent/assent form home with him/her to review it
with family/friends.� The subject will
be told that his/her participation in the study is voluntary and will be
reassured that s/he may elect not to participate without affecting his/her
relationship with UCLA or with the investigator.� The subject will be told that his/her participation will in no
way affect the relationship with the physician or the treatment that the
subject will receive from the physician.�
The alternatives to participating in the study will be reviewed with the
subject.� The consent /assent will be
obtained in a private room.
24. � Comprehension
of the Information Provided:� How--and
by whom--will it be determined whether the subjects or their legally authorized
representatives understand the information provided?
������� Note: This section should clearly document
that the investigator has an adequate plan in place to assure existence of an
acceptable level of comprehension before consent is documented.� The principal investigator (or approved
designee) is responsible for assuring that prospective subjects or their
representatives have sufficient understanding of the research to make an
informed decision about participation.�
It is important that they understand the purpose of the research, the
nature and duration of the procedures, any risks and discomforts involved, the
possible benefits to the subjects and others, and their right to withdraw
consent at any time without penalty.�
Willingness to sign the consent form is not an adequate demonstration of
their understanding.� Some investigators
try to determine the level of prospective subjects� comprehension by
questioning them about the research.�
(This approach is useful with children and adolescents, as well as with
adults of uncertain capacity to consent.)
The potential subject will be asked by the
investigator whether s/he understood the information in the consent/assent
form. The potential subject will be asked questions about information in the
consent/assent form to ensure understanding.�
The investigator will review information in the consent form with the
subject to ensure comprehension of the requirements of the study and of each
study visit.� The subject will be
questioned to ensure that s/he understands the risks associated with the study,
the potential benefits, and the alternatives to participation in the study.
Parents/guardians will be asked the same type of questions.
25. � Information Withheld From Subjects:� Will any information about the research
purpose and design be withheld from potential or participating subjects?� if so, explain and justify the
non-disclosure and describe plans for post-study debriefing.
������� Note:�
Any non-disclosure must be approved by the HSPC and may not exclude
information that a reasonable person would want to know in deciding whether to
participate in the research.� In
addition, the alteration in the consent procedure must be approvable under 45
CFR 46.116(d): (1) the research involves no more than minimal risk to the
subjects; (2) the waiver or alteration will not adversely affect the rights and
welfare of the subjects; (3) the research could not practicably be carried out
without the waiver or alternation; and (4) whenever appropriate, the subjects
will be provided with additional pertinent information after participation.
�������
No information will be withheld
26. � Consent/Assent Forms:� Specify the form(s) that will be used
among the following: adult consent form, parental consent form, proxy consent
form, youth assent form (age 13-18), and/or child assent form (age 7-12).
�������
Subjects 18 years of age and older will use the adult
consent form.� Subjects 13-17 will use
the youth assent form.�
Parents/guardians of subjects 13-17 will be given the adult consent
form.