Human T-lymphocyte response in vitro to synthetic peptides of herpes simplex virus glycoprotein D.

Abstract

Immunization of mice with a synthetic peptide that corresponds to a murine antibody-defined immunodominant domain of herpes simplex virus (HSV) glycoprotein D (gD) induced neutralizing antibodies against HSV types 1 and 2 and protected animals against a lethal challenge with HSV type 2 (Dietzschold, B., Eisenberg, R., Ponce de Leon, M., Golub, E., Hudecz, F., Varicchio, A. & Cohen, G. (1984) J. Virol. 52,[431][432][433][434][435]. We report here that human peripheral blood T cells from HSV-seropositive and -seronegative adult donors are activated by this synthetic peptide in vitro. Interleukin-2-dependent T-cell lines established from these cultures respond specifically to peptides containing residues 1-23 of HSV gD and to a panel of overlapping peptides within this domain. The T-cell proliferative response was maximal when the majority of interleukin-2-propagated T cells were of the helper phenotype and the peptides were at least 16 amino acids long. Peptides of 8 or 12 amino acids from the carboxyl terminus were nonstimulatory. Peptide-activated T-cell lines from seronegative donors less than 11 years old could be established in vitro, but most cells were of the suppressor/cytotoxic phenotype and demonstrated no antigen-specificity when tested with the panel of synthetic peptides.The immune response to herpes simplex virus (HSV) has been shown to involve a variety of effector mechanisms which include production of virus-neutralizing antibody, cytotoxic T lymphocytes, natural killer (NK) cells, antibodydependent cellular cytotoxicity, and T-cell-derived lymphokines (43). The pivotal role of helper T-cells in the induction of these responses has been well documented (44,45).The nature of the HSV-specified antigens that act as immunostimulants has not been completely identified, but several lines of evidence implicate the major viral glycoproteins (1). One of these, glycoprotein D (gD), has been shown to induce type-common viral-neutralizing antibody (2, 3), and passive administration of monoclonal antibody (mAb) specific for gD protects mice from a lethal challenge with HSV (4). Moreover, gD appears to be recognized on HSV-infected cells by cytolytic effector cells (5).An antibody-defined immunodominant determinant of HSV type 1 (HSV-1) gD was localized and a peptide corresponding to residues 8-23 of the mature glycoprotein was synthesized. This peptide was recognized by an anti-gD mAb known to neutralize HSV-1 and -2 (6). Moreover, immunization of mice with this synthetic molecule induces viral-neutralizing antibodies and confers protection against a lethal challenge with HSV-2 (7). MATERIALS AND METHODSSynthesis of HSV gD Peptides. Peptides were manually synthesized as described (6) by using the Merrifield solidphase method (8). The completed peptides were simultaneously deprotected and cleaved from the resin with anhydrous HF containing thioanisole as an organic cation scavenger. The crude peptides were purified by gel filtration on Bio-Gel P-2 or P-4 (Bio-Rad) columns. Homogeneity of the peptides w...

Gender-Dependent HLA-DR-Restricted Epitopes Identified from Herpes Simplex Virus Type 1 Glycoprotein D

“…Moreover, the present strategy accurately identified regions bearing all of the previously reported gD epitopes: gD (24,28,36), gD [241][242][243][244][245][246][247][248][249][250][251][252][253][254][255][256][257][258][259][260] (34,48,80), and gD 290-314 (24). Therefore, our approach to epitope identification, as well as testing epitope peptides for their T-cell antigenicity, immunogenicity, and protective efficacy, should be nearly universal, regardless of the protein studied.…”

Identification of Novel Immunodominant CD4+Th1-Type T-Cell Peptide Epitopes from Herpes Simplex Virus Glycoprotein D That Confer Protective Immunity

“…The latter epitopes were identified with other methods (e.g., overlapping peptides and truncated forms of gD protein) and other algorithms (e.g., prediction of amphipathic and hydrophilic parameters) (10,23,35,36,50,63,68,78). The present strategy accurately identified regions bearing all of the previously reported gD epitopes: gD (13,20,22,36); gD [241][242][243][244][245][246][247][248][249][250][251][252][253][254][255][256][257][258][259][260] (35,53,83), and gD (20). More importantly, the present approach identified several novel, not previously described, Th1 immunodominant CD4 ϩ T-cell epitopes that elicited protective immunity.…”

“…Of the 11 known HSV glycoproteins, gD is the most highly conserved and the most antigenically cross-reactive between HSV-1 and HSV-2 (61). gD has emerged as an excellent candidate Ag for inducing a protective immunity in animal models against ocular and genital infections with both types of HSV (22,35,36,50,54,63,73,80). It has recently been reported in a large human genital HSV trial that immunization with recombinant gD induced significant protection in women who were seronegative for HSV but was ineffective in seropositive women and in men (41,74).…”

“…The gD epitopes and the immune mechanism that provide protective immunity are not yet fully understood. Knowledge of immune responses to gD and of their correlation with protection is limited primarily to the identification of B-cell epitopes (15,18,73,80), with only a limited number of T-cell epitopes having been reported to date (13,20,22,35,36,53,74,83). We hypothesized that characterization of the CD4 ϩ T-cell epitope repertoire of gD could be vital in the case of HSV infection.…”

Regulation of interleukin 2 receptors on T cells from multiple sclerosis patients.

“…All donors used had been immunized previously for medical reasons with tetanus toxoid and bacillus Calmette-Guerin (BCG Clones of antigen-induced lines were established by limiting dilution in the presence of autologous AC, homologous antigen, and rIL-2 (50 ng/ml). Antigen-induced T-cell lines and clones were tested for specificity by proliferation as described (9).…”