Table 1 Characteristics of peptide and protein vaccine studies

Arp, J

1993

USA

in vivo

BALB/c (Charles River), C57BL/6 (CharlesRiver) and CFW/D

envelope protein inclusion bodies (env-I.B.) in the presence or absence of an adjuvant

10 µg

IP

10 µg of env-I.B at 6 and 8 weeks of age then boosed 2 and 4 weeks later

Western blotting, radioimmunoprecipitation, peptide ELISA and a syncytium inhibition assay

Antibodies against the HTLV-I env protein in the presence or absence of an adjuvant, neutralizing Ab in admission of high doses of mycobacterial cell wall extract, enhanced Ab response to the HTLV-I envelope glycoprotein following priming with recombinant vaccinia virus (Rvv) constructs expressing either the entire native HTLV-I envelope (gp46 and gp21) or just gp46, Increased titres of neutralizing Ab following priming with the Rvv expressing gp46 only

Baba, E

1995

Japan

in vivo

Female New Zealand White rabbits and inbred female WWQdj, Fisher 433 (F433)/Qdj rats, Inbred female BN/Sea, LewisiSea rats, d ACI/Jcl rats

2 vaccines: T and B cell epitope-based peptide vaccine constructed from the conjugation of gp46 (aa 181–210) and (181–203) with a branched polylysine oligomer

500 µg (rabbit)

100 µg (rat)

IM/SC

9 New Zealand White rabbits were immunized 500 µg of pKA- or OVA-conjugated peptide (MAP181-203, MAP181- 210, 181-2030VA) emulsified with CFA on day 0 and then with IFA on days 14 and 28

5 different inbred strains of rats similarly immunized either i.m. or S.C. with 100 pg of pKA-conjugated peptide (MAP181-203, MAP181-210) emulsified with CFA on day 0 and then with IFA on days 14 and 28. Control rabbits and rats were immunized with the same amounts of CFA and IFA only

western blotting, ELISA, PCR, IF

Neutralizing Ab production in rabbits ( X4 -8 and X8-64 titers in response to MAPl 81–203 and MAPl 81 -21 0, respectively), neutralizing Abs (X40 to X320) in five different strains of rats in response to MAP1 81–210

Lairmore

1995

USA

In vivo

"Female inbred strains of mice (BALB/c, C3H/ HeJ, and C57BL/6) were obtained from Jackson Laboratories (Bar Harbor, Maine), and outbred ICR mice were obtained from Harlan Industries (Indianapolis, Ind.). Rabbits "

Chimeric and b-template peptide constructs incorporating known human T-lymphotropic virus type 1 (HTLV-1) B- and T-cell epitopes from the surface envelope protein gp46 (SP2 [aa 86 to 107] and SP4a [aa 190 to 209]) and promiscuous T-cell peptides were synthesized

100 µg in mice/ 500 µg in rabbits

SC

Booster in 3 weeks in mice/ 2 weeks in rabbits

Competitive ELISA, The radioimmunoprecipitation assay, A human osteosarcoma cell-based assay

Promiscuous T-cell epitopes, which bind to several forms of human MHC class II molecules, can be used with immunodominant peptides derived from retroviruses to produce highly immunogenic response

Schönbach

1996

Japan

in vitro and in vivo

Eight- to ten-week-old transgenic HLA-B-3501 transgenic mice of both sexes

synthetic HTLV-1 peptides mixed with the lipohexapeptide N-palmitoyl-S-[2,3-bis(palmitoyloxy)propyl]cysteinyl-seryl-lysyl-lysyl-lysyl-lysine, which is a biocompatible, Thepitopeindependent adjuvant

100 µM

intraperitoneally

NA

Peptide binding assay, Flow cytometric analysis, Western blot analysis, Cytotoxic T lymphocyte assay

CTL response in response to 11 of 37 tested HLA-B-3501 binding peptides after 3 in vitro stimulations, peptide-specific CTL induction in response to 7 peptides derived from env-gp46 (VPSPSSTPLL, VPSSSSTPL, YPSLALAPH, and YPSLALAPA), pol (QAFPQCTIL), gagp19 (YPGRVNEIL), and tax (GAFLTNVPY) proteins, Bulk CTL generation by four peptides derived from env-gp46 (SPPSTPLLY, VPSPSSTPLLY, and VPSPSSTPLL) and pol (QAFPQCTILQY) killing peptide-pulsed and recombinant vaccinia-infected target cells

Hanabuchi, S

2001

Japan

in vivo

Four-week-old female F344/N Jcl-rnu/rnu (nu/nu or athymic) rats and F344/N Jcl-rnu/ + (nu/ +) rats

synthetic oligopeptides corresponding to the Tax-epitope(180–188)

100 microg Tax 180–188 peptide alone, 10 nmol of ISS-ODN alone, 100 microg Tax 180–188 peptide mixed with 10 nmol of ISS ODN (Tax 180–188/ISS-ODN), or 100 microg Influenza A matrix 58–66 peptide mixed with 10 nmol of ISS-ODN (Influenza A matrix 58–66/ISS-ODN)

ID/IP

twice with a 2-week interval

Two weeks after the last immunization, 107 freshly isolated T-cell enriched splenocytes from vaccinated rats were intraperitoneally inoculated into 4-week old nu/nu rats, which were simultaneously inoculated subcutaneously with FPM1-V1AX cells

cold inhibition assay/Cr-release assay/peptide mapping

Development prevention of FPM1-V1AX cell induced lymphomas in athymic rats in response to adoptive transfer of the Tax 180–188-specific CTL line or freshly prepared T cells from rats vaccinated with the Tax 180–188 oligopeptide in comparison with control groups, equivalent inhibitory effects on the growth of HTLV-I-infected tumors in both CD4 + and CD8 + T cells, Tax 180–188 as a dominant epitope recognized by the HTLV-I Tax-specific CTL line

Frangione-Beebe, M

2001

USA

in vivo

female New Zealand white rabbits (12 weeks)

vaccine (MVFMF2) comprising HTLV-1-gp46 (aa 175–218) linked by GPSL turn to MVF (aa 288–302)

Two rabbits: 18 mg of microspheres containing 1 mg of peptide, and 3.7 mg of microspheres containing 100 microg of adjuvant (nor-MDP)

Two additional rabbits: 18 mg microspheres containing 1 mg of peptide and no adjuvant. Microspheres containing peptide or adjuvant were suspended in 1 ml of 4:1 squalene-arlacel A (Sigma) and injected intramuscularly remaining two rabbits: 1 mg of free peptide (1.3 mg/mL in PBS) and 100 mg of nor-MDP emulsied 50:50 in 4:1 squalene: arlacel A + boosting with 500 mg of peptide and 100 mg of nor-MDP at 10 weeks

IM

4 rabbits once in 12 weeks

2 rabbits in 10 and 12 weeks

ELISA, HPLC, Gun scanning electron microscope,

Sustained antibody response over a period of 5 months, without requiring a booster immunization or adjuvant in response to encapsulation of MVFMF2, elevated immune response invoked by the encapsulated peptide without requiring booster and adjuvant, Raised Ab level against both free and encapsulated MVFMF2

Frangione-Beebe, M

2000

USA

in vivo

Outbred female ICR mice female New Zealand white rabbits

MVFMF2 comprising HTLV-1-gp46 (aa 175–218) linked by GPSL turn to MVF (aa 288–302)

1 mg of peptide

IM

different for host groups

PCR, SIA, ELISA, western blot, circular dichroism (CD) spectroscopy

enhanced reactivity to viral antigens in rabbits, high titered anti-peptide antibodies in mice, immunogenic in an outbred population of both rabbits and mice when administered with adjuvant, enhanced immunogenicity when encapsulated in biodegradable microspheres without requiring of adjuvant, syncytium formation inhibition ability of anti-rabbit and anti-mouse Abs, no protection from cell-associated viral challenge in rabbits

Sundaram, R

2003

USA

in vivo

transgenic HHD mice

three HLA-A ∗ 0201 restricted CTL epitopes (Tax11–19, Tax178–186, and Tax233–241)

100 g of peptide mixed with 140 g TT3

SC

twice, 3 weeks apart

ELISPOT, Cr release assay

cellular responses to each intended epitope in vivo, high level of IFN-γ production

Sundaram, R

2004

USA

in vivo

leukocyte antigen–A-0201 monochain trans genic H-2Db -2 m double-knockout (HHD) mice

multivalent cytotoxic T-lymphocyte peptide construct derived from the Tax protein ofHTLV-1 separated by arginine spacers

100 µg of multiepitope peptide or with a mixture of 33 µg of each of the 3 individual epitopes combined with 140 µg of TT3, a pro miscuous T-helper epitope from tetanus toxoid (residues 947–967) and 100 µg of adjuvant N-acetyl-glucosamine-3-acetyl L-alanyl-D-isoglutamine (nor-MDP; Peninsula Laboratories, Belmont, CA)22 emulsified 50:50 in 4:1 squalene/Arlacel A (Sigma, St. Louis, MO)

SC

twice, 3 weeks apart

reverse-phase high-performance liquid chromatography (RP-HPLC), Cr Release Assay, interferon-gamma Release Assay, ELISA, Plaque Assay for Viral Titers

significant reduction in viral replication dependent on CD8 + T cells

Sundaram

2004

USA

in vivo

Female New Zealand outbred white rabbits, female ICR mice (8 weeks)

chimeric synthetic B-cell epitopes derived from HTLV-1-Env (gp21 and gp46) with promiscuous T-helper epitopes derived either from the tetanus toxoid (amino acids 947–967) or measles virus fusion protein (amino acids 288–302)

rabbits: initial dose (1 mg of the chimeric peptide) + booster (500 µg of peptide) / mice: initial dose (100 µg of peptide) + booster (500 µg of peptide)

s.c. in the thigh muscle in 2 rabbits, s.c in mice

rabbits [initial dose + booster injections every 3–5 weeks apart], mice [initial dose + booster at 3 and 6 weeks]

Circular dichroism spectroscopy, computer-aided analyses of protein antigenicity, Syncytia inhibition assay, Flow cytometry, Immunogenicity testing,

Neutralizing Ab against the epitopes derived from the gp21, inhibition the formation of virus-induced syncytia, peptid had secondary structure correlated well with the crystal structure data or predicted structure

Kazanji, M

2006

France

in vivo

male squirrel monkeys

three HLA-A-0201-restricted CTL epitopes derived from Tax protein (Tri-Tax) and B-cell env epitope (aa 175–218)

700 microg

IM

Two monkeys were injected twice, at 0 and 4 weeks, with the Env B-cell epitope aa 175–218 (500 mg per monkey) linked to the promiscuous T-helper cell epitope MVF (700 mg per monkey), as described pre viously (Frangione-Beebe et al., 2000). Six weeks after the first immunization, the monkeys were injected with another construct, consisting of the three Tax CTL epitopes (aa 11–19, 178–186, 306–315). Monkeys were boosted twice at weeks 9and 16 with both B- and T-cell epitopes

ELISA, PCR, western blot

high titre of Abs, high frequency of specific IFN-c-producing cells and partial protection

Kobayashi

2006

Florida

in vitro

Cell lines: EBV-LCLs, Mouse fibroblast cell lines (L-cells), HTLV-I-infected T-cell lymphoma cell lines TL-Su, TCL-Kan, HUT102, TL-Hir (HTLV-1 Tax negative), and OKM-2 T, Jurkat T-cell lymphoma cell line (HTLV-I negative), MT2,

Potential HLA-DR-restricted CD4 + T-cell epitopes of HTLV-1 Tax peptid

NA

NA

NA

Western blot analysis, ELISA, ECL detection system, chemiluminescence assay, cytokine release assay, high-performance liquid chromatography, mass spectrometry,

T-helper-cell induction in response to peptides Tax191–205 (restricted by the HLA-DR1 and DR9 alleles) and Tax305–319 (restricted by either DR15 or DQ9), Both these epitopes were naturally processed by HTLV-1 + T-cell lymphoma cells and by autologous APCs that were pulsed with HTLV-1Tax + tumor lysates. These epitopes lie proximal to known CTL epitopes, which will facilitate the development of prophylactic peptide – based vaccine capable of inducing simultaneous CTL andT-helper responses

Kozako, T

2009

Japan

in vivo

HLA-A-0201-transgenic mice

HTLV-1/hepatitis B virus core (HBc) chimeric particle incorporating the HLA-A-0201-restricted HTLV1 Tax-epitope

HTLV-1/HBc chimeric particle (20microg), or Tax11–19 peptide / HBc particle (20microg) and peptide (1microg)

intradermally

days 0 and 14 with HTLV-1/HBc chimeric particle (20 microg), or Tax11–19 peptide

ELISPOT, PCR, western blot, FCA, ELISA, enzyme-linked immunospot assay

induction of HTLV-1 Tax-specific CD8 + cells from spleen and inguinal lymph nodes after immunization, efficient induction of IFN- -producing cells, antigen-specific gamma-interferon reaction induction, increased expression of CD86, HLA-A02, TLR4 and MHC class II in dendritic cells, HTLV-1-specific CD8 + T-cells induction by peptide with HTLV-1/HBc particle from ATL patient, but not by peptide only, lysing cell presenting the peptide by HTLV-1-specific CD8 + T-cells

Kozako, T

2011

Japan

in vivo

HLA-A-0201-transgenic mice

an HTLV-1-specific CD8 + T-cell response by oligomannose-coated liposomes (OMLs) encapsulating the human leukocyte antigen (HLA)A-0201-restricted HTLV-1 Tax-epitope (OML⁄Tax)

1 microg

IM

days 0 and 14 with OML⁄Tax, Tax peptide alone or phosphate-buffered saline (PBS)

ELISPOT, FCM assay

resulted in the efficient induction of IFN-gamma-producing cells, induction of HTLV-1 Tax-specific CD8 + cells from inguinal lymph nodesafter immunization with OML/Tax, increased CD86,MHCI, HLA-A02 and MHCII levels upon exposure of dendritic cells to OML⁄Tax

Kuo, C. W

2011

Scotland

NA

NA

gp46

soluble recombinant surface glycoprotein (gp46, SU) fused to the Fc region of human IgG (sRgp46-Fc)

NA

NA

ELISA, Western blot, Syncytium interference assay, Flow cytometry,

High titer Ab responses/ Many of these mAbs recognize envelope displayed on the surface of HTLV-1–infected cells / mAbs robustly antagonize envelope-mediated membrane fusion and neutralize pseudovirus infectivity/ Potent neutralizing mAbs recognize the N-terminal receptor-binding domain / Both neutralizing and poorly neutralizing Abs strongly stimulate neutrophil-mediated cytotoxic responses to HTLV-1–infected cells

Kobayashi

2012

Florida

in vitro

Cell lines: EBV-LCLs, Mouse fibroblast cell lines (L cells), HTLV-1 infected T cell lymphoma cell lines, TL-Su, TCL-Kan,OKM-2 T, Hut102, TL-Hir, Jurkat T cell lymphoma cell line, prostate tumor cell line PC3, and PBMCs

HLA-DR-bound peptide from the IL-9 receptor alpha of HTLV-1-transformed T cells

NA

NA

NA

Purification of HLA-DR molecules, Preparation of bound peptides, Binding assay, Western blot analyses, Cell-mediated cytotoxicity assays,

antigen-specific CD4 helper T lymphocytes generation (in vitro) restricted by HLA-DR15 or HLA-DR53 molecules with recognizing and killing ability of HTLV-1 + , IL-9Ra + T cell lymphoma cells

Fujisawa

2015

Japan

in vivo

HTLV-1-infected humanized mouse model (hu-NOG) mouce

Mixture of twelve overlapping peptides of 40–42 amino acids long encompassing whole Tax protein

NA

subcutaneously

vaccine was inoculated subcutaneously three times weekly to hu-NOG mouse and then -irradiated HTLV-1 producing Jurkat cells were intraperitoneally injected to infect HTLV-1

NA

leukemia suppression, retardation of the out growth of human lymphocytes in response to Tax-immunization after HTLV-1 infection, survival of two out of five mice with alimited number of infected T-cells, IL-12 induction and enhanced expression of Tax-specific CD8 T-cell in immunized mice before infection

Amirnasr, M

2016

Iran

in vivo

male BALB/c mice

env23 (162–209) and env13 (125–209) recombinant proteins

7.5 µg antigen

nasal/SC

3 immunizations (7.5 µg antigen) were performed with 2 weeks intervals

ELISA, PCR

higher serum IgG1 and IgG total levels compared to antigen solution, higher IgG2a levels and IgG2a/IgG1 ratio in nasal delivery compared with subcutaneous administration (P < 0.001), higher cellular immune responses in response to env23 antigen, compared with env13

Kabiri, M

2018

Iran

in vivo

BALB/c male mice

chimeric peptide vaccine including Tax, gp21, gp46, and gag immunodominant epitopes of human T-cell lymphotropic virus type 1 (HTLV-1)

10 microg

nasal/SC

three times at two weeks intervals

ELISA, PCR

increased Ab titers containing IgG2a, mucosal IgA, as well as IFN-γ and IL-10 cytokines and decreased TGF-β1 level in response to mixture of IMX and chimera, potent mucosal sIgA titers in intranasal delivery compared to subcutaneous root, cell-mediated responses, as evident by higher IgG2a and IFN-γ, as well as suppressed TGF-β1 level in SC or nasal delivery

Kabiri, M

2018

Iran

in vivo

BALB/c male mice

chimeric peptide vaccine including Tax (aa 11–19 and aa 178–186), gp21 (aa 370–400), gp46 (aa 165–306), and p19 (aa 105–124) immunodominant epitopes of human T-cell lymphotropic virus type 1 (HTLV-1)

10 microg

nasal/SC

three times at two weeks intervals

ELISA, PCR, western blot

elevated titers of IgG1, IgG2a, and sIgA antibodies, as well as IL-10, and IFN-γ cytokines and decreased TGF-β1 level, promoted cellular and mucosal responses in co-delivery of chimera and CpG ODN in PLGA

Mulherkar

2018

USA

in vivo and in vitro

Six to eight-week old female HLA-A2 transgenic mice / Cell lines for in vitro investigations: HepG2, hepatoma cells, MT2, HTLV-1 virion expressing cells, and T2, TAP deficient lymphoblasts

MHC-I-bound HTLV-1 peptides

NA

interadermal near the base of the tail and subcutaneous on the flank

Three injections: initial inoculation (consisted of a mixture of pooled free peptide in PBS plus Montanide ISA 51 (Seppic, Paris, France) (50:50 emulsion), PBS alone, or two independent, individual free peptide in PBS plus Montanide ISA 51 (50:50 emulsion)) + repeated two more times at 10-day intervals)

Degranulation assay, CD8 + T-cell killing assay, Mass spectrometry analysis, Flow cytometry analysis, ELISpot assays, MagPix cytokine detection,

confirmation of six novel MHC-I restricted epitopes capable of binding HLA-A2 and HLA-A24 alleles, generation of CD8 + T cells specific for each of these peptides, generation of epitope-specific CD8 + T cells secreted IFN-γ, granzyme B, MIP-1α, TNF-α, perforin and IL-10 in the presence of MT-2 cell line in vitro, cytotoxic response through surface expression of CD107 on CD8 + T cells when cultured with MT-2 cells, significant antiviral activity of CD8 + T cells specific against all identified peptides, In vivo generation of CD8 + T cells similarly demonstrated immunogenicity on ELISpot, CD107 degranulation assay, and MagPix MILLIPLEX analysis

Pandey

2019

India

in silico

NA

vaccine by the assimilation of B‐cell, CTL, and HTL epitopes for GAG, POL, ENV, P12, P13, P30, REX, and TAX proteins

NA

NA

NA

B‐cell, Helper T‐cell (HTL), and Cytotoxic T‐cell (CTL) epitope prediction, Tertiary structure prediction, Molecular docking, in silico cloning

interactions with the HLA-A0201, HLA-A0701 and HLA-A0301 receptors, Strong interaction with TLR-3

Alam

2020

Bangladesh?

in silico

NA

prediction of 14 epitopes for targeting Glycoprotein 62

NA

NA

NA

Variability Analysis of GP62 of HTLV-1, Population Protection Coverage (PPC) Calculation, HLA-Epitope Binding Prediction, Molecular Dynamics Simulation, Prediction ofB-Cell Epitope,

ALQTGITLV and VPSSSTPL epitopes interaction with three MHC alleles ( including HLA-A-02:03, and HLA-B-35:01, respectively), 70% summative population protection coverage

Jahantigh

2021

Iran

in silico

NA

eight-epitopes-rich domain, including overlapping epitopes detected on both B and T cells constructed of Gag, Env, Pol, Hbz, and Tax proteins

NA

NA

NA

antigen prediction, Mapping, 3D Structure modeling, Homology modeling, Antigenicity and allergenicity and solubility and other physicochemical parameters evaluation, structure prediction, In silico cloning, Immune simulation, peptide–allele docking

interaction of the epitope and the designed protein with immune receptors(in silico docking), strong interaction of O2 epitope and D8 protein with immune receptors especially the HLAA 02:01 receptor, stability of the interactions for 100 ns(molecular dynamic), root mean square deviation, radius of gyration, hydrogen bonds, and solvent-accessible surface area were calculated for the 100 ns, humoral and cell-mediated immune responses elicited

Raza

2021

Bangladesh

in silico

NA

predicting HTLV-1 TAX multiepitope protein constructed from CTL and B cell epitopes

NA

NA

NA

Primary, secondary, tertiary and 3-D structure analysis, B- and T-cell epitope prediction, molecular docking analysis, Disulfide engineering, in silico cloning,

most antigenic score of 0.57, strong T cell epitopes interaction with HLA-A-0201, high binding affinity of the vaccine construct for TLR4 (in molecular docking study), most antigenic and immunogenic epitopes in in-silico investigation: B cell epitopes (KEADDNDHEPQISPGGLEPPSEKHFR and DGTPMISGPCPKDGQPS spanning from 324–349 and 252–268 respectively); T cell epitopes (LLFGYPVYV, ITWPLLPHV and GLLPFHSTL ranging from 11–19, 163–171 and 233–241)

Tariq

2021

Pakistan

in silico

NA

9 Cytotoxic T Lymphocytes, 6 Helper T Lymphocytes and 5 Linear B Lymphocytes epitopes, joint through linkers and adjuvant

NA

NA

NA

Conservation analysis and selection of predicted epitopes, Epitope modeling and molecular docking, homology analysis, Disulphide engineering, In-silico estimation and cloning,

strong binding affinity with their corresponding Human Leukocyte Antigen alleles, 95.8% coverage of the world’s population, highly antigenic properties while being non-toxic, soluble, non-allergenic, and stable in nature, enhanced stability via disulphide engineering, strong association between vaccine construct and human pathogenic immune receptor TLR3 (in Molecular docking analysis and Molecular Dynamics (MD)), rapid antigen clearance and higher levels of cell-mediated immunity in response to repeated-exposure and immune simulations, respectively

shafifar

2022

Iran

in vivo and in vitro

male 6 to 8 weeks pathogen-free BALB/c mice

Fc-fusion multi-immunodominant recombinant protein (Tax-Env: mFcγ2a and Tax-Env: His)

1) Six mice received 50 μg of tTax-tEnv:mFcγ2a in 100 μL PBS + 100 μL of DDA adjuvant /2) Four mice, 50 μg of tTax-tEnv:His in 100 μL of PBS + 100 μL of DDA adjuvant / 3) Five mice, 200 μL of PBS (negative control)

intraperitoneal

T200 μL/mouse thrice at two-week intervals (0, 14th, and 28th days)

SDS-PAGE, Western blot, real time PCR

significant increase in IFN-γ and IL-12 release in response to Tax-Env: mFcγ2a compared to Tax-Env: His, 50% low proviral load of HTLV-1 and 50% complete protection in challenged mice, more Th1 immune responses in response to "Tax-Env: mFcγ2a”, more Th2 immune responses in response to “Tax-Env: His”