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HIV-1 induces cytotoxic T lymphocytes in the cervix of infected women.

Musey L, Hu Y, Eckert L, Christensen M, Karchmer T, McElrath MJ - J. Exp. Med. (1997)

Bottom Line: Class II MHC-restricted CD4+ CTL clones lysed targets expressing Env gp41 or infected with HIV-1.Class I MHC-restricted CD8+ clones recognized HIV-1 Gag- or Pol-expressing targets, and the epitopes were mapped to within 9-20 amino acids.Comparisons of intra-individual cervical and blood CTL specificities indicate that epitopes recognized by CTL in the cervix were commonly recognized in the blood.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, The University of Washington School of Medicine, Seattle 98195, USA.

ABSTRACT
Although T lymphocytes are present in the genital mucosa, their function in sexually transmitted diseases is unproven. To determine if cervical T cells mediate HIV-specific cytolysis, mononuclear cells in cytobrush specimens from HIV-1-infected women were stimulated in vitro with antigen. Resultant cell lines lysed autologous targets expressing HIV-1 proteins in 12/19 (63%) subjects, and these responses were detected intermittently on repeated visits. All 8 subjects with blood CD4+ counts > or =500 cells/microl had HIV-1-specific cervical CTL, whereas only 4/11 with counts <500 cells/microl had detectable responses (P = 0.008). Class II MHC-restricted CD4+ CTL clones lysed targets expressing Env gp41 or infected with HIV-1. Class I MHC-restricted CD8+ clones recognized HIV-1 Gag- or Pol-expressing targets, and the epitopes were mapped to within 9-20 amino acids. Comparisons of intra-individual cervical and blood CTL specificities indicate that epitopes recognized by CTL in the cervix were commonly recognized in the blood. These studies provide the first definitive evidence for an MHC-restricted effector function in human cervical lymphocytes.

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Cervical CTL clones derived from subject 738 (visit 8) are  restricted by HLA-A2 and recognize targets expressing HIV-1 Pol aa 476484 or HIV-1-infected cells. (A) Specific lysis of autologous and MHC  class I partially matched allogeneic B-LCL infected with either vRT or  the control, vSC-8, at an E/T 3:1. Serological HLA class I typing for the  donors are the following: autologous, (A2,74, B58,63, Bw4,w6); allo-1,  (A2,28,B44,14,Bw4,w6); allo-2, (A2,B27,44,Bw4); allo-3, (A23,74,B70,57,  Bw4,w6). (B) Lysis by clones of autologous B-LCL pulsed with synthetic  peptides spanning Pol amino acid region 346-354 (VIYQYMDDL) and  476-484 (ILKEPVHGV) at an E/T of 3:1. C. Lysis at E/T of 3:1 of HIV1-infected and uninfected autologous B-LCL. Targets were infected with  either HIV-1LAI or one of two primary patient isolates, no. 1002 or no.  1005.
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Figure 4: Cervical CTL clones derived from subject 738 (visit 8) are restricted by HLA-A2 and recognize targets expressing HIV-1 Pol aa 476484 or HIV-1-infected cells. (A) Specific lysis of autologous and MHC class I partially matched allogeneic B-LCL infected with either vRT or the control, vSC-8, at an E/T 3:1. Serological HLA class I typing for the donors are the following: autologous, (A2,74, B58,63, Bw4,w6); allo-1, (A2,28,B44,14,Bw4,w6); allo-2, (A2,B27,44,Bw4); allo-3, (A23,74,B70,57, Bw4,w6). (B) Lysis by clones of autologous B-LCL pulsed with synthetic peptides spanning Pol amino acid region 346-354 (VIYQYMDDL) and 476-484 (ILKEPVHGV) at an E/T of 3:1. C. Lysis at E/T of 3:1 of HIV1-infected and uninfected autologous B-LCL. Targets were infected with either HIV-1LAI or one of two primary patient isolates, no. 1002 or no. 1005.

Mentions: In donor 738, four CD8+ clones were derived from the cervical cell line at visit 8 (Fig. 2). These clones recognized and lysed targets expressing HIV-1 Pol with a specific lysis ranging from 4161% at an E/T of 3:1 (Fig. 4 A). To determine the MHC restriction patterns of these CTL clones, cytolysis of vRTinfected B-LCL partially matched at MHC class I loci was compared to vRT-infected autologous B-LCL. As shown in Fig. 4 A, lysis of the Pol-specific target cells was restricted to HLA-A2, with comparable lysis of targets from donors 115 (allo-1) and 5224 (allo-2) who share this haplotype. To identify the epitope recognized by the MHCrestricted CTL clones, we tested two nonamer peptides previously reported to be associated with HLA-A2-restricted HIV-1 Pol CTL (24). The 4 clones lysed autologous targets pulsed with the synthetic peptide spanning aa 476-484 but not the peptide spanning aa 346-354 of the Pol region (Fig. 4 B). To ascertain the ability of these CTL clones to lyse not only target cells expressing Pol through infection with recombinant vaccinia or pulsing with peptides, but also infected with HIV-1, we tested 4 clones against autologous B-LCL targets infected with either HIV-1LAI or one of two primary HIV-1 isolates obtained from PBMC co-cultures of recently infected patients. As shown in Fig. 4 C, lysis by the four clones was observed at an E/T of 3:1 against targets infected with the laboratory-adapted strain HIV-1LAI (49, 38, 39, and 22%), but not against targets infected with the two primary isolates (specific lysis <1%) or against the control uninfected targets (lysis ⩽1%). In summary, these experiments demonstrate that the HLA-A2-restricted cervical CD8+CTL clones derived from this subject recognize a known epitope within HIV-1 Pol characteristic of peripheral blood CTL, and that the mucosal CD8+ CTL, in contrast to the mucosal CD4+ CTL, may function to destroy some but not all HIV-1 strains.


HIV-1 induces cytotoxic T lymphocytes in the cervix of infected women.

Musey L, Hu Y, Eckert L, Christensen M, Karchmer T, McElrath MJ - J. Exp. Med. (1997)

Cervical CTL clones derived from subject 738 (visit 8) are  restricted by HLA-A2 and recognize targets expressing HIV-1 Pol aa 476484 or HIV-1-infected cells. (A) Specific lysis of autologous and MHC  class I partially matched allogeneic B-LCL infected with either vRT or  the control, vSC-8, at an E/T 3:1. Serological HLA class I typing for the  donors are the following: autologous, (A2,74, B58,63, Bw4,w6); allo-1,  (A2,28,B44,14,Bw4,w6); allo-2, (A2,B27,44,Bw4); allo-3, (A23,74,B70,57,  Bw4,w6). (B) Lysis by clones of autologous B-LCL pulsed with synthetic  peptides spanning Pol amino acid region 346-354 (VIYQYMDDL) and  476-484 (ILKEPVHGV) at an E/T of 3:1. C. Lysis at E/T of 3:1 of HIV1-infected and uninfected autologous B-LCL. Targets were infected with  either HIV-1LAI or one of two primary patient isolates, no. 1002 or no.  1005.
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Related In: Results  -  Collection

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Figure 4: Cervical CTL clones derived from subject 738 (visit 8) are restricted by HLA-A2 and recognize targets expressing HIV-1 Pol aa 476484 or HIV-1-infected cells. (A) Specific lysis of autologous and MHC class I partially matched allogeneic B-LCL infected with either vRT or the control, vSC-8, at an E/T 3:1. Serological HLA class I typing for the donors are the following: autologous, (A2,74, B58,63, Bw4,w6); allo-1, (A2,28,B44,14,Bw4,w6); allo-2, (A2,B27,44,Bw4); allo-3, (A23,74,B70,57, Bw4,w6). (B) Lysis by clones of autologous B-LCL pulsed with synthetic peptides spanning Pol amino acid region 346-354 (VIYQYMDDL) and 476-484 (ILKEPVHGV) at an E/T of 3:1. C. Lysis at E/T of 3:1 of HIV1-infected and uninfected autologous B-LCL. Targets were infected with either HIV-1LAI or one of two primary patient isolates, no. 1002 or no. 1005.
Mentions: In donor 738, four CD8+ clones were derived from the cervical cell line at visit 8 (Fig. 2). These clones recognized and lysed targets expressing HIV-1 Pol with a specific lysis ranging from 4161% at an E/T of 3:1 (Fig. 4 A). To determine the MHC restriction patterns of these CTL clones, cytolysis of vRTinfected B-LCL partially matched at MHC class I loci was compared to vRT-infected autologous B-LCL. As shown in Fig. 4 A, lysis of the Pol-specific target cells was restricted to HLA-A2, with comparable lysis of targets from donors 115 (allo-1) and 5224 (allo-2) who share this haplotype. To identify the epitope recognized by the MHCrestricted CTL clones, we tested two nonamer peptides previously reported to be associated with HLA-A2-restricted HIV-1 Pol CTL (24). The 4 clones lysed autologous targets pulsed with the synthetic peptide spanning aa 476-484 but not the peptide spanning aa 346-354 of the Pol region (Fig. 4 B). To ascertain the ability of these CTL clones to lyse not only target cells expressing Pol through infection with recombinant vaccinia or pulsing with peptides, but also infected with HIV-1, we tested 4 clones against autologous B-LCL targets infected with either HIV-1LAI or one of two primary HIV-1 isolates obtained from PBMC co-cultures of recently infected patients. As shown in Fig. 4 C, lysis by the four clones was observed at an E/T of 3:1 against targets infected with the laboratory-adapted strain HIV-1LAI (49, 38, 39, and 22%), but not against targets infected with the two primary isolates (specific lysis <1%) or against the control uninfected targets (lysis ⩽1%). In summary, these experiments demonstrate that the HLA-A2-restricted cervical CD8+CTL clones derived from this subject recognize a known epitope within HIV-1 Pol characteristic of peripheral blood CTL, and that the mucosal CD8+ CTL, in contrast to the mucosal CD4+ CTL, may function to destroy some but not all HIV-1 strains.

Bottom Line: Class II MHC-restricted CD4+ CTL clones lysed targets expressing Env gp41 or infected with HIV-1.Class I MHC-restricted CD8+ clones recognized HIV-1 Gag- or Pol-expressing targets, and the epitopes were mapped to within 9-20 amino acids.Comparisons of intra-individual cervical and blood CTL specificities indicate that epitopes recognized by CTL in the cervix were commonly recognized in the blood.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, The University of Washington School of Medicine, Seattle 98195, USA.

ABSTRACT
Although T lymphocytes are present in the genital mucosa, their function in sexually transmitted diseases is unproven. To determine if cervical T cells mediate HIV-specific cytolysis, mononuclear cells in cytobrush specimens from HIV-1-infected women were stimulated in vitro with antigen. Resultant cell lines lysed autologous targets expressing HIV-1 proteins in 12/19 (63%) subjects, and these responses were detected intermittently on repeated visits. All 8 subjects with blood CD4+ counts > or =500 cells/microl had HIV-1-specific cervical CTL, whereas only 4/11 with counts <500 cells/microl had detectable responses (P = 0.008). Class II MHC-restricted CD4+ CTL clones lysed targets expressing Env gp41 or infected with HIV-1. Class I MHC-restricted CD8+ clones recognized HIV-1 Gag- or Pol-expressing targets, and the epitopes were mapped to within 9-20 amino acids. Comparisons of intra-individual cervical and blood CTL specificities indicate that epitopes recognized by CTL in the cervix were commonly recognized in the blood. These studies provide the first definitive evidence for an MHC-restricted effector function in human cervical lymphocytes.

Show MeSH
Related in: MedlinePlus