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Spermatozoa capture HIV-1 through heparan sulfate and efficiently transmit the virus to dendritic cells.

Ceballos A, Remes Lenicov F, Sabatté J, Rodríguez Rodrígues C, Cabrini M, Jancic C, Raiden S, Donaldson M, Agustín Pasqualini R, Marin-Briggiler C, Vazquez-Levin M, Capani F, Amigorena S, Geffner J - J. Exp. Med. (2009)

Bottom Line: Interaction of spermatozoa with DCs not only leads to the transmission of HIV-1 and the internalization of the spermatozoa but also results in the phenotypic maturation of DCs and the production of IL-10 but not IL-12p70.At low values of extracellular pH (approximately 6.5 pH units), similar to those found in the vaginal mucosa after sexual intercourse, the binding of HIV-1 to the spermatozoa and the consequent transmission of HIV-1 to DCs were strongly enhanced.Our observations support the notion that far from being a passive carrier, spermatozoa acting in concert with DCs might affect the early course of sexual transmission of HIV-1 infection.

View Article: PubMed Central - HTML - PubMed

Affiliation: Centro Nacional de Referencia para SIDA, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires C1121ABG, Argentina.

ABSTRACT
Semen is the main vector for HIV-1 dissemination worldwide. It contains three major sources of infectious virus: free virions, infected leukocytes, and spermatozoa-associated virions. We focused on the interaction of HIV-1 with human spermatozoa and dendritic cells (DCs). We report that heparan sulfate is expressed in spermatozoa and plays an important role in the capture of HIV-1. Spermatozoa-attached virus is efficiently transmitted to DCs, macrophages, and T cells. Interaction of spermatozoa with DCs not only leads to the transmission of HIV-1 and the internalization of the spermatozoa but also results in the phenotypic maturation of DCs and the production of IL-10 but not IL-12p70. At low values of extracellular pH (approximately 6.5 pH units), similar to those found in the vaginal mucosa after sexual intercourse, the binding of HIV-1 to the spermatozoa and the consequent transmission of HIV-1 to DCs were strongly enhanced. Our observations support the notion that far from being a passive carrier, spermatozoa acting in concert with DCs might affect the early course of sexual transmission of HIV-1 infection.

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MRs do not play a major role in the capture of HIV-1 by spermatozoa. (A) Spermatozoa were treated with 1,000 U/ml trypsin or 20 µg/ml pronase for 15 min at 37°C, washed thoroughly, and 1.5 × 106/200 µl were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C. Cells were then washed, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of five experiments performed in triplicate. Asterisks represent statistical significance (P < 0.05 vs. controls). (B) The expression of MRs in the spermatozoa was analyzed by measuring the binding of albumin bovine-α-d-mannopyranosylphenyl isothiocyanate-BMA revealed by streptavidin-FITC. Assays were performed by incubating spermatozoa for 30 min at 37°C with 100 µg/ml BMA in the absence or presence of 5 mg/ml mannan. The gray histogram represents spermatozoa incubated only with streptavidin-FITC (isotype). A representative experiment (n = 4) is shown. (C) Spermatozoa (1.5 × 106/200 µl) were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan, mannose-BSA, or blocking antibodies directed to MR. Cells were washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of four to eight experiments performed in duplicate. (D) Human macrophages (105/100 µl), obtained from monocytes cultured with GM-CSF for 5 d, were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan, mannose-BSA, or blocking antibodies directed to MR. Cells were washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of three to four experiments performed in duplicate. (E) B-THP-1-DC-SIGN+ cells (2.5 × 105/200 µl) were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan or mannose-BSA, washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of three to five experiments performed in duplicate. Asterisks in C–E represent statistical significance (P < 0.05 vs. controls). The capture mediated by THP-1 cells, which do not express DC-SIGN, is also shown (black bar).
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fig2: MRs do not play a major role in the capture of HIV-1 by spermatozoa. (A) Spermatozoa were treated with 1,000 U/ml trypsin or 20 µg/ml pronase for 15 min at 37°C, washed thoroughly, and 1.5 × 106/200 µl were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C. Cells were then washed, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of five experiments performed in triplicate. Asterisks represent statistical significance (P < 0.05 vs. controls). (B) The expression of MRs in the spermatozoa was analyzed by measuring the binding of albumin bovine-α-d-mannopyranosylphenyl isothiocyanate-BMA revealed by streptavidin-FITC. Assays were performed by incubating spermatozoa for 30 min at 37°C with 100 µg/ml BMA in the absence or presence of 5 mg/ml mannan. The gray histogram represents spermatozoa incubated only with streptavidin-FITC (isotype). A representative experiment (n = 4) is shown. (C) Spermatozoa (1.5 × 106/200 µl) were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan, mannose-BSA, or blocking antibodies directed to MR. Cells were washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of four to eight experiments performed in duplicate. (D) Human macrophages (105/100 µl), obtained from monocytes cultured with GM-CSF for 5 d, were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan, mannose-BSA, or blocking antibodies directed to MR. Cells were washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of three to four experiments performed in duplicate. (E) B-THP-1-DC-SIGN+ cells (2.5 × 105/200 µl) were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan or mannose-BSA, washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of three to five experiments performed in duplicate. Asterisks in C–E represent statistical significance (P < 0.05 vs. controls). The capture mediated by THP-1 cells, which do not express DC-SIGN, is also shown (black bar).

Mentions: Fig. 2 A shows that pretreatment of spermatozoa with either trypsin or pronase almost completely prevented the binding of HIV-1, suggesting that a protein receptor mediates HIV binding. We then analyzed the expression of the different known receptors for HIV-1 in human spermatozoa. Consistent with previously published data (el-Demiry et al., 1986; Wolff and Anderson, 1988; Kim et al., 1999), CD4 expression was not detected on the spermatozoa (unpublished data). Other published studies have shown that spermatozoa express MR (Benoff et al., 1993; Chen et al., 1995). Consistent with these studies, we found that spermatozoa bind biotin-labeled mannose coupled to albumin (BMA) and that the binding of BMA to spermatozoa was almost completely inhibited by mannan, an inhibitor of the family of C-type lectin receptors, including MR and DC-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN; Hong et al., 2002; Nguyen and Hildreth, 2003; Fig. 2 B). Treatment of spermatozoa with 1,000 U/ml trypsin for 15 min at 37°C resulted in a diminished ability to bind BMA, with a 79 ± 18% decrease in the mean fluorescence intensity (MFI; mean ± SEM; n = 4; P < 0.01, untreated vs. trypsin-treated spermatozoa).


Spermatozoa capture HIV-1 through heparan sulfate and efficiently transmit the virus to dendritic cells.

Ceballos A, Remes Lenicov F, Sabatté J, Rodríguez Rodrígues C, Cabrini M, Jancic C, Raiden S, Donaldson M, Agustín Pasqualini R, Marin-Briggiler C, Vazquez-Levin M, Capani F, Amigorena S, Geffner J - J. Exp. Med. (2009)

MRs do not play a major role in the capture of HIV-1 by spermatozoa. (A) Spermatozoa were treated with 1,000 U/ml trypsin or 20 µg/ml pronase for 15 min at 37°C, washed thoroughly, and 1.5 × 106/200 µl were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C. Cells were then washed, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of five experiments performed in triplicate. Asterisks represent statistical significance (P < 0.05 vs. controls). (B) The expression of MRs in the spermatozoa was analyzed by measuring the binding of albumin bovine-α-d-mannopyranosylphenyl isothiocyanate-BMA revealed by streptavidin-FITC. Assays were performed by incubating spermatozoa for 30 min at 37°C with 100 µg/ml BMA in the absence or presence of 5 mg/ml mannan. The gray histogram represents spermatozoa incubated only with streptavidin-FITC (isotype). A representative experiment (n = 4) is shown. (C) Spermatozoa (1.5 × 106/200 µl) were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan, mannose-BSA, or blocking antibodies directed to MR. Cells were washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of four to eight experiments performed in duplicate. (D) Human macrophages (105/100 µl), obtained from monocytes cultured with GM-CSF for 5 d, were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan, mannose-BSA, or blocking antibodies directed to MR. Cells were washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of three to four experiments performed in duplicate. (E) B-THP-1-DC-SIGN+ cells (2.5 × 105/200 µl) were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan or mannose-BSA, washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of three to five experiments performed in duplicate. Asterisks in C–E represent statistical significance (P < 0.05 vs. controls). The capture mediated by THP-1 cells, which do not express DC-SIGN, is also shown (black bar).
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Related In: Results  -  Collection

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fig2: MRs do not play a major role in the capture of HIV-1 by spermatozoa. (A) Spermatozoa were treated with 1,000 U/ml trypsin or 20 µg/ml pronase for 15 min at 37°C, washed thoroughly, and 1.5 × 106/200 µl were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C. Cells were then washed, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of five experiments performed in triplicate. Asterisks represent statistical significance (P < 0.05 vs. controls). (B) The expression of MRs in the spermatozoa was analyzed by measuring the binding of albumin bovine-α-d-mannopyranosylphenyl isothiocyanate-BMA revealed by streptavidin-FITC. Assays were performed by incubating spermatozoa for 30 min at 37°C with 100 µg/ml BMA in the absence or presence of 5 mg/ml mannan. The gray histogram represents spermatozoa incubated only with streptavidin-FITC (isotype). A representative experiment (n = 4) is shown. (C) Spermatozoa (1.5 × 106/200 µl) were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan, mannose-BSA, or blocking antibodies directed to MR. Cells were washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of four to eight experiments performed in duplicate. (D) Human macrophages (105/100 µl), obtained from monocytes cultured with GM-CSF for 5 d, were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan, mannose-BSA, or blocking antibodies directed to MR. Cells were washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of three to four experiments performed in duplicate. (E) B-THP-1-DC-SIGN+ cells (2.5 × 105/200 µl) were incubated with HIV-1 BAL containing 25 ng p24 for 60 min at 37°C in the absence or presence of mannan or mannose-BSA, washed thoroughly, lysed, and assayed for p24 antigen by ELISA. Results are the mean ± SEM of three to five experiments performed in duplicate. Asterisks in C–E represent statistical significance (P < 0.05 vs. controls). The capture mediated by THP-1 cells, which do not express DC-SIGN, is also shown (black bar).
Mentions: Fig. 2 A shows that pretreatment of spermatozoa with either trypsin or pronase almost completely prevented the binding of HIV-1, suggesting that a protein receptor mediates HIV binding. We then analyzed the expression of the different known receptors for HIV-1 in human spermatozoa. Consistent with previously published data (el-Demiry et al., 1986; Wolff and Anderson, 1988; Kim et al., 1999), CD4 expression was not detected on the spermatozoa (unpublished data). Other published studies have shown that spermatozoa express MR (Benoff et al., 1993; Chen et al., 1995). Consistent with these studies, we found that spermatozoa bind biotin-labeled mannose coupled to albumin (BMA) and that the binding of BMA to spermatozoa was almost completely inhibited by mannan, an inhibitor of the family of C-type lectin receptors, including MR and DC-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN; Hong et al., 2002; Nguyen and Hildreth, 2003; Fig. 2 B). Treatment of spermatozoa with 1,000 U/ml trypsin for 15 min at 37°C resulted in a diminished ability to bind BMA, with a 79 ± 18% decrease in the mean fluorescence intensity (MFI; mean ± SEM; n = 4; P < 0.01, untreated vs. trypsin-treated spermatozoa).

Bottom Line: Interaction of spermatozoa with DCs not only leads to the transmission of HIV-1 and the internalization of the spermatozoa but also results in the phenotypic maturation of DCs and the production of IL-10 but not IL-12p70.At low values of extracellular pH (approximately 6.5 pH units), similar to those found in the vaginal mucosa after sexual intercourse, the binding of HIV-1 to the spermatozoa and the consequent transmission of HIV-1 to DCs were strongly enhanced.Our observations support the notion that far from being a passive carrier, spermatozoa acting in concert with DCs might affect the early course of sexual transmission of HIV-1 infection.

View Article: PubMed Central - HTML - PubMed

Affiliation: Centro Nacional de Referencia para SIDA, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires C1121ABG, Argentina.

ABSTRACT
Semen is the main vector for HIV-1 dissemination worldwide. It contains three major sources of infectious virus: free virions, infected leukocytes, and spermatozoa-associated virions. We focused on the interaction of HIV-1 with human spermatozoa and dendritic cells (DCs). We report that heparan sulfate is expressed in spermatozoa and plays an important role in the capture of HIV-1. Spermatozoa-attached virus is efficiently transmitted to DCs, macrophages, and T cells. Interaction of spermatozoa with DCs not only leads to the transmission of HIV-1 and the internalization of the spermatozoa but also results in the phenotypic maturation of DCs and the production of IL-10 but not IL-12p70. At low values of extracellular pH (approximately 6.5 pH units), similar to those found in the vaginal mucosa after sexual intercourse, the binding of HIV-1 to the spermatozoa and the consequent transmission of HIV-1 to DCs were strongly enhanced. Our observations support the notion that far from being a passive carrier, spermatozoa acting in concert with DCs might affect the early course of sexual transmission of HIV-1 infection.

Show MeSH
Related in: MedlinePlus