Expression of multi-copy and immuno-reactivity of recombinant type-specific epitope of herpes simplex virus type 1 / 中华男科学杂志

<p><b>OBJECTIVE</b>To express a multi-copy specific epitope recombinant protein of herpes simplex virus type 1 (HSV-1) with immuno-reactivity.</p><p><b>METHODS</b>Multi-copy genes with a specific epitope of HSV-1-glycoprotein G 112-127 were constructed by DNA recombination and cloned in E. coli JM109 pGEM-5Zf. The positive recombinants were determined by SDS-PAGE and Western blotting.</p><p><b>RESULTS</b>The recombinants with 4, 8, 16 and 32 copies of gG 112-127 were obtained. The 8-copy recombinant was expressed by 17.5%, mainly as inclusion body. And it reacted with antiserum HSV-1, but not with antiserum HSV-2.</p><p><b>CONCLUSION</b>The HSV-1-gG112-127 recombinant could be used to distinguish HSV-1 and HSV-2 in ELISA.</p>

CTL epitopes modified by KDEL and recognized by CD8+ T lymphocytes to herpes simplex virus type 2 improve CTL effect / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the improvement of CTL effect by CTL epitopes which are modified by KDEL and recognized by CD8+ T lymphocytes to herpes simplex virus type 2.</p><p><b>METHODS</b>Observations were made on the specific immune response induced by the CD8+ CTL epitopes(SSIEFARL, S1), the CD8+ CTL epitopes modified by KDEL(SSIEFARL-KDEL, S1-KDEL), the tandem four copies CD8+ CTL epitopes[(SSIEFARL)4, S4] and the tandem four copies CD8+ CTL epitopes modified by KDEL[(SSIEFARL)4-KDEL, S4-KDEL], 25 male C57BL/6 mouse were randomly divided into 5 group, 5 mice per group, respectively immunized with istonic Na chloride, S1, S1-KDEL, S4 and S4-KDEL. Lymphocyte proliferation was detected by 3H-TdR and the CTL effect induced by CTL epitopes in vivo was detected by 51Cr.</p><p><b>RESULTS</b>In the 3H-TdR test, compared with the control, the group S1 and group S1-KDEL, the cpm values of Group S4 and Group S4-KDEL were markedly higher (P < 0.05) and the cpm value of Group S4-KDEL was significantly higher than Group S4 (P < 0.05), but the cpm values of Group S1 and Group S1-KDEL were not significantly different from the control (P > 0.05), nor was that of Group S1 from Group S1-KDEL (P > 0.05). In the experiment in which the EL4 cells sensitized by S1 were attacked as target cells, the CTL activities induced in Group S4 and Group S4-KDEL were markedly higher (P < 0.05) compared with the control, Group S1 and Group S1-KDEL, and that induced in Group S4-KDEL was significantly higher than Group S4 (P < 0.05), but the CTL activity induced in Group S1 and Group S1-KDEL were not significantly different from the control (P > 0.05), nor was that induced in Group S1 from Group S1-KDEL (P > 0.05). In the experiment in which the EL4 cells were attacked as target cells, the kill rate was below 10% in every group, not significantly different from the control.</p><p><b>CONCLUSION</b>The tandem four copies CD8+ CTL epitopes, modified by KDEL and recognized by CD8+ T lymphocytes to herpes simplex virus type 2, can improve the CTL effect.</p>