M A J O R A R T I C L E
Specificity and 6-Month Durability of Immune
Responses Induced by DNA and Recombinant
Modified Vaccinia Ankara Vaccines Expressing
HIV-1 Virus-Like Particles
Paul A. Goepfert,1,aMarnie L. Elizaga,2,aKelly Seaton,4Georgia D. Tomaras,4David C. Montefiori,4Alicia Sato,2
John Hural,2Stephen C. DeRosa,2,3Spyros A. Kalams,5M. Juliana McElrath,2,3Michael C. Keefer,6Lindsey R. Baden,9
Javier R. Lama,15Jorge Sanchez,15Mark J. Mulligan,10Susan P. Buchbinder,12Scott M. Hammer,7Beryl A. Koblin,8
Michael Pensiero,13Chris Butler,13Bernard Moss,14and Harriet L. Robinson,11for the HVTN 205 Study Group,band the
National Institutes of Allergy and Infectious Diseases HIV Vaccines Trials Network
1Department of Medicine, University of Alabama at Birmingham;2Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, and
3University of Washington, Seattle, Washington;4Laboratory for AIDS Vaccine Research and Development, Department of Surgery, Duke University
Medical Center, Durham, North Carolina;5Vanderbilt University School of Medicine, Nashville, Tennessee;6University of Rochester School of Medicine
and Dentistry, Rochester, and7Columbia Universityand8New York Blood Center, New York, New York;9Brigham and Women’s Hospital, Harvard Medical
School, Boston, Massachusetts;10Division of Infectious Diseases, Emory University, Atlanta, and11GeoVax Inc., Smyrna, Georgia;12Bridge HIV,
San Francisco Department of Public Health, California;13Division of AIDS and14Laboratory of Viral Diseases, National Institute of Allergy and Infectious
Diseases, National Institutes of Health, Bethesda, Maryland; and15Asociacion Civil IMPACTA Salud y Educacion, Lima, Peru
ticles displaying trimeric membrane-bound envelope glycoprotein (Env) were tested in a phase 2a trial in human immu-
nodeficiency virus (HIV)–uninfected adults for safety, immunogenicity, and 6-month durability of immune responses.
months, respectively (the DDMM regimen); 3 doses of MVA/HIV62B at 0, 2, and 6 months (the MMM regimen); or
Results. At peak response, 93.2% of the DDMM group and 98.4% of the MMM group had binding antibodies for
Env. These binding antibodies were more frequent and of higher magnitude for the transmembrane subunit (gp41)
than the receptor-binding subunit (gp120) of Env. For both regimens, response rates were higher for CD4+T cells
(66.4% in the DDMM group and 43.1% in the MMM group) than for CD8+T cells (21.8% in the DDMM group
and 14.9% in the MMM group). Responding CD4+and CD8+T cells were biased toward Gag, and >70% produced
2 or3 ofthe4 cytokines evaluated (ie,interferonγ, interleukin 2, tumornecrosisfactorα, and granzyme B). Six months
after vaccination, the magnitudes of antibodies and T-cell responses had decreased by <3-fold.
Conclusions.DDMM and MMM vaccinations with virus-like particle–expressing immunogens elicited durable
antibody and T-cell responses.
Clade B DNA and recombinant modified vaccinia Ankara (MVA) vaccines producing virus-like par-
Keywords.HIV/AIDS; vaccines; clinical trial; T cells; antibodies; DNA; recombinant MVA.
A human immunodeficiency virus (HIV) vaccine faces
the challenge of eliciting immune responses that can
prevent the acquisition of virus and the establishment
of latency. Vaccine-induced antibodies (Abs) can
block infection by directly neutralizing virus  and
by binding to virus and virus-infected cells to tag
them for destruction by the innate immune response
[2,3]. Elicited cytotoxic (ie, CD8+) T cells can modulate
the severity of infection and slow disease progression by
recognizing and killing infected cells.
Received 4 October 2013; accepted 19 December 2013; electronically published 7
Presented in part: AIDS Vaccine Meeting, Boston, Massachusetts, September
2012. Abstract 0A09.08 LB.
aP. A. G. and M. L. E. contributed equally to this work.
bAdditional members of the study group are listed at the end of the text.
Correspondence: Paul A. Goepfert, MD, 908 20th St South, CCB 328, Birmingham,
AL 35294 (firstname.lastname@example.org).
The Journal of Infectious Diseases 2014;210:99–110
© The Author 2014. Published by Oxford University Press on behalf of the Infectious
Diseases Society of America. All rights reserved. For Permissions, please e-mail:
HIV Vaccine–Induced Durable Abs • JID 2014:210 (1 July) • 99
by guest on July 10, 2015
The one vaccine to achieve at least partial prevention of infec-
tion (efficacy, 31.2%) was tested in Thailand in the RV144 trial
. This vaccine regimen consisted of 2 recombinant canarypox
primes(ALVAC-HIVvCP1521),followedby2 canarypox plusbi-
valent gp120 protein in alum boosts (AIDSVAX B/E) . The
elicited Abs had limited ability to neutralize tier 1 HIV isolates,
which are easy to neutralize, and no detectable neutralizing activ-
ity for tier 2 viruses, which are more difficult to neutralize and
characteristic of most currently circulating viruses .In the cor-
circulating Env-specific immunoglobulin A (IgA) appeared to de-
crease vaccine efficacy by competing with immunoglobulin G
(IgG) binding and Fcγ-initiated mechanisms of protection, such
as Ab-dependent cellular cytotoxicity [7,10,11].Nonneutralizing
Ab, induced by the RV144 vaccine regimen, captured infectious
RV144 rapidly waned, fallingby ≥10-fold in the first 6 months [6,
13]. Efficacy also fell with time, from an estimated 60% at peak
vaccine response to <30% by 2.5 years .
The DNA and recombinant modified vaccinia Ankara
(MVA62B) vaccines tested in this phase 2a study produce
virus-like particles that display membrane-bound trimeric
forms of Env [15–17]. Simian immunodeficiency virus (SIV)
prototypes of these vaccines elicited 61%–64% reductions in
the per-challenge risk of intrarectal infection and prevented in-
fection in 25% of the animals receiving 12 weekly rectal admin-
istrations of the heterologous SIVE660 . Phase 1 testing of
these vaccines revealed that both the DDMM regimen, involv-
ing 2 doses of DNA vaccine followed by 2 doses of MVA62B
vaccine, and the MMM regimen, involving 3 doses of
MVA62B vaccine, were well tolerated and replicated the overall
patterns of immunogenicity observed for analogous SIV immu-
nogens in macaques [19,20]. The current phase 2atrial expand-
ed testing of the DDMM and MMM regimens and extended
analyses of both T-cell and Ab responses to include the durabil-
ity of elicited responses at 6 months after vaccination.
SUBJECTS, MATERIALS, AND METHODS
The GeoVax vaccine, GOVX-B11 comprises a DNA prime and
B HIV-HXB-2/BH10 sequences and Env from HIV ADA
sequences (gp41/120 cleavage site intact) from a single transcript
by subgenomic splicing . The vaccine is rendered nonin-
fectious by gene deletions and inactivating point mutations [15,
16]. The MVA component, MVA62B, encodes HIV-1 Gag, pro-
tease, reverse transcriptase, and Env from the same sequences
and also produces immature noninfectious virus-like particles
. In MVA62B, the ADA Env gene is truncated for the 115
C-terminal amino acids of the endodomain of gp41 to enhance
stability of the vaccine insert during manufacture .
HIV Vaccine Trials Network (HVTN) protocol 205 was a ran-
domized, double-blind, placebo-controlled trial conducted at
clinical sites in the United States and Peru among participants
who were considered to be at lower risk for HIV infection (clin-
ical trials registration NCT00820846). The institutional review
boards or ethics committees for each site provided initial and
ongoing approvals and review of the research. Adults aged
18–50 years who were deemed healthy on the basis of medical
history, physical examination findings, laboratory test results,
troponin levels, and electrocardiogram (ECG) findings were en-
rolled. In part A of the study, 180 participants were enrolled, of
whom 120 were vaccinated with 3 mg of DNA at months 0 and
2, followed by 108median tissue culture infective doses of
MVA62B at months 4 and 6 (the DDMM regimen); 60 addi-
tional participants were enrolled and received normal saline as
placebo injections. In part B, 29 enrolled participants received
DDMM, and 75 enrolled participants received MVA62B at
months 0, 2, and 6 (the MMM regimen); 15 additional enrolled
subjects received normal saline as placebo injections.
Vaccines were delivered intramuscularly by needle injection at
a final volume of 1 mL into the deltoid region. Safety evaluations
included physical examinations, standard clinical chemistry and
hematological tests, and cardiac troponin analysis. Postvaccina-
tion chest symptoms were evaluated with a 12-lead ECG, and
findings were interpreted bya central ECG laboratory. Local reac-
togenicity (ie, injection site pain, tenderness, redness, erythema,
and induration) and systemic reactogenicity (ie, malaise, head-
ache, fever, chills, myalgias, arthralgias, nausea, vomiting, and fa-
tigue) were assessed for3 days followingeachvaccination or until
resolution. Adverse events were recorded for 12 months after
the first vaccination for each participant and were graded as
mild, moderate, or severe according to standard criteria (avail-
able at: http://rcc.tech-res.com/safetyandpharmacovigilance/).
Social impact assessments were obtained at each study visit fol-
lowing the first vaccination and consisted of 10 targeted ques-
tions about potential discrimination due to study participation.
Immune Response Assays
Validated binding Ab multiplex assays  for IgG and IgA
were performed according to a prespecified assay study plan
and good clinical laboratory practices guidelines. HIV-specific
anti-IgG Abs were detected with mouse anti-human IgG
(Southern Biotech, Birmingham, AL). Anti-HIV IgA responses
in serum were detected with goat anti-human IgA (Jackson Im-
munoresearch, West Grove, PA) in specimens depleted of IgG
by use of protein G high-performance MultiTrap plates (GE
100 • JID 2014:210 (1 July) • Goepfert et al
by guest on July 10, 2015
STUDY GROUP MEMBERS
In addition to authors of this article, members of the HVTN
205 Study Group consist of Yeycy Donastorg, Li Qin, Dale Law-
rence, Massimo Cardinali, Jin Bae, Renée Holt, Huguette Red-
inger, Jan Johannessen, Gail Broder, Jerri Moody-White, Butch
McKay, Gabriela Calazans, Carter Bentley, Lisa Kakinami, Katie
Skibinski, Scharla Estep, Jenny Tseng, Molly Swenson, Tamra
Madenwald, Edgar Turner Overton, Srilatha Edupuganti, Na-
dine Rouphael, Jennifer Whitaker, C Mhorag Hay, Catherine
A Bunce, Pedro Gonzales, Juan Carlos Hurtado, Raphael
Dolin, Ken Mayer, Steven Walsh, and Jennifer Johnson.
Supplementary materials are available at The Journal of Infectious Diseases
online (http://jid.oxfordjournals.org/). Supplementary materials consist of
data provided by the author that are published to benefit the reader. The
posted materials are not copyedited. The contents of all supplementary
dataare thesole responsibilityofthe authors. Questions ormessagesregarding
errors should be addressed to the author.
bers at the HVTN research clinics.
Health (NIH; grants AI69452 [to the University of Alabama–Birmingham],
AI069418 [to Emory University], AI06970 [to the New York Blood Center
and Columbia University Medical Center], AI069511 [to the University of
Rochester], AI069438 [to the Asociacion Civil Impacta Salud y Educacion],
AI069412 [to Brigham and Women’s Hospital], AI069439 [to Vanderbilt
University], AI069481 [to the Fred Hutchinson Cancer Research Center],
AI069496 [to the San Francisco Department of Public Health], AI068614
[to the HVTN Core] AI068635 [to SCHARP], and AI068618 [to the
HVTN Laboratory]) and the NIH Integrated Preclinical/Clinical AIDS Vac-
cine Development Program (grant U19A1074073 to GeoVax for product
and preclinical studies).
Potential conflict of interest.
GeoVax. M. J. M. works for Emory, which is a stakeholder in GeoVax,
but M. J. M. does not have any direct holdings in GeoVax. All other authors
report no potential conflicts.
All authors have submitted the ICMJE Form for Disclosure of Potential
Conflicts of Interest. Conflicts that the editors consider relevant to the
content of the manuscript have been disclosed.
We thank the study participants and the staff mem-
H. L. R. is a stakeholder in
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