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Infection of semen-producing organs by SIV during the acute and chronic stages of the disease.

Le Tortorec A, Le Grand R, Denis H, Satie AP, Mannioui K, Roques P, Maillard A, Daniels S, Jégou B, Dejucq-Rainsford N - PLoS ONE (2008)

Bottom Line: We demonstrate for the first time the presence of SIV in the testes, epididymides, prostate and seminal vesicles as early as 14 days post-inoculation.The prostate and seminal vesicles appear to be the most efficiently infected reproductive organs, followed by the epididymides and testes.Within the male genital tract, mostly T lymphocytes and a small number of germ cells harbour SIV antigens and RNA.

View Article: PubMed Central - PubMed

Affiliation: INSERM U625, Rennes, University of Rennes I, Groupe d'Etude de la Reproduction chez l'Homme et les Mammifères, IFR 140, Campus de Beaulieu, Rennes, France.

ABSTRACT

Background: Although indirect evidence suggests the male genital tract as a possible source of persistent HIV shedding in semen during antiretroviral therapy, this phenomenon is poorly understood due to the difficulty of sampling semen-producing organs in HIV+ asymptomatic individuals.

Methodology/principal findings: Using a range of molecular and cell biological techniques, this study investigates SIV infection within reproductive organs of macaques during the acute and chronic stages of the disease. We demonstrate for the first time the presence of SIV in the testes, epididymides, prostate and seminal vesicles as early as 14 days post-inoculation. This infection persists throughout the chronic stage and positively correlates with blood viremia. The prostate and seminal vesicles appear to be the most efficiently infected reproductive organs, followed by the epididymides and testes. Within the male genital tract, mostly T lymphocytes and a small number of germ cells harbour SIV antigens and RNA. In contrast to the other organs studied, the testis does not display an immune response to the infection. Testosteronemia is transiently increased during the early phase of the infection but spermatogenesis remains unaffected.

Conclusions/significance: The present study reveals that SIV infection of the macaque male genital tract is an early event and that semen-producing organs display differential infection levels and immune responses. These results help elucidate the origin of HIV in semen and constitute an essential base to improving the design of antiretroviral therapies to eradicate virus from semen.

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Related in: MedlinePlus

SIV localization within the male genital tract.Detection of SIV positive cells in the seminal vesicles (A, C), testes (B–B', D–D'), epididymides (E, E') and prostate (F) of primary-infected macaques using immunohistochemistry for SIVp27 (A, B) and in situ hybridization (ISH) for SIV gag RNA (C–F). The phenotype of SIV positive cells was determined using ISH for SIV gag RNA (visualized as black silver grains) combined with immunostaining for cell markers (visualized as brown staining): combined ISH for viral RNA and immunostaining for CD3 revealed black silver grains clustered over brown cells in the seminal vesicles (C), prostate (F), and epididymis (E'), indicating infection of CD3+T lymphocytes. Co-labelling of SIV RNA+ cells with the myeloid cell marker CD68 was also observed, as shown here for the epididymis (E). In the testis, SIV RNA was detected within the interstitium in HLA-DR+ cells (D) and within the seminiferous tubules in VASA+ germ cells (D'). Inserts show enlargement of SIV RNA positive cells co-stained for cell markers. I: testicular interstitium; ST: seminiferous tubules. Scale bars = 20 µm. (G) SIV RNA+ cells were counted in a minimum of 30 tissue sections/experiment in 3 independent experiments on a primary-infected macaque MGT. Results show the mean positive cell number/tissue area +/− SEM.
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pone-0001792-g004: SIV localization within the male genital tract.Detection of SIV positive cells in the seminal vesicles (A, C), testes (B–B', D–D'), epididymides (E, E') and prostate (F) of primary-infected macaques using immunohistochemistry for SIVp27 (A, B) and in situ hybridization (ISH) for SIV gag RNA (C–F). The phenotype of SIV positive cells was determined using ISH for SIV gag RNA (visualized as black silver grains) combined with immunostaining for cell markers (visualized as brown staining): combined ISH for viral RNA and immunostaining for CD3 revealed black silver grains clustered over brown cells in the seminal vesicles (C), prostate (F), and epididymis (E'), indicating infection of CD3+T lymphocytes. Co-labelling of SIV RNA+ cells with the myeloid cell marker CD68 was also observed, as shown here for the epididymis (E). In the testis, SIV RNA was detected within the interstitium in HLA-DR+ cells (D) and within the seminiferous tubules in VASA+ germ cells (D'). Inserts show enlargement of SIV RNA positive cells co-stained for cell markers. I: testicular interstitium; ST: seminiferous tubules. Scale bars = 20 µm. (G) SIV RNA+ cells were counted in a minimum of 30 tissue sections/experiment in 3 independent experiments on a primary-infected macaque MGT. Results show the mean positive cell number/tissue area +/− SEM.

Mentions: SIV p27 positive cells were found within the stroma and close to the secretory epithelium of the seminal vesicles (Figure 4A), prostate and epididymides (data not shown) of primary-infected animals. Positive staining was observed in the interstitium and seminiferous tubules of the testis (Figure 4B, B'). In situ hybridization for SIV gag RNA also revealed positive cells in the stroma and, occasionally, within the epithelium of the seminal vesicles (Figure 4C), epididymides (Figure 4E, E') and prostate (Figure 4F) of primary-infected animals. Within the testis, SIV RNA+cells were observed in the interstitium (Figure 4D), occasionally bordering the seminiferous tubules (data not shown). In all tissues, these infected cells were mainly T lymphocytes (60–97% of SIV RNA+ cells co-labeled with CD3, the highest proportion being found in the prostate and seminal vesicles) (Figure 4C, E', F), and some macrophages (0–25% of cells co-labeled with CD68, the highest rate being consistently found in the epididymis) (Figure 4E). Interestingly, in the testis SIV gag positive cells that never co-localized with either HLA-DR, CD3 or CD68 were occasionally observed within the seminiferous tubules (Figure 4D') (on average 1 positive cell for 300 seminiferous tubules). These positive cells systematically co-localized with VASA, a specific germ cell marker [42]. Their distribution and localization, from the base to the middle of the tubules, suggested an association of SIV with pre-meiotic and meiotic germ cells. In chronically infected macaques, the same pattern of SIV localization was observed, but fewer cells were affected. Importantly, however, a few SIV+ cells were detected within the MGT of an animal with undetectable blood viremia (data not shown). A quantitative measurement of the number of SIV+ cells per tissue surface in a primary-infected animal evidenced that seminal vesicles and prostate displayed a higher number of positive cells when compared to the epididymides and testes (Figure 4G).


Infection of semen-producing organs by SIV during the acute and chronic stages of the disease.

Le Tortorec A, Le Grand R, Denis H, Satie AP, Mannioui K, Roques P, Maillard A, Daniels S, Jégou B, Dejucq-Rainsford N - PLoS ONE (2008)

SIV localization within the male genital tract.Detection of SIV positive cells in the seminal vesicles (A, C), testes (B–B', D–D'), epididymides (E, E') and prostate (F) of primary-infected macaques using immunohistochemistry for SIVp27 (A, B) and in situ hybridization (ISH) for SIV gag RNA (C–F). The phenotype of SIV positive cells was determined using ISH for SIV gag RNA (visualized as black silver grains) combined with immunostaining for cell markers (visualized as brown staining): combined ISH for viral RNA and immunostaining for CD3 revealed black silver grains clustered over brown cells in the seminal vesicles (C), prostate (F), and epididymis (E'), indicating infection of CD3+T lymphocytes. Co-labelling of SIV RNA+ cells with the myeloid cell marker CD68 was also observed, as shown here for the epididymis (E). In the testis, SIV RNA was detected within the interstitium in HLA-DR+ cells (D) and within the seminiferous tubules in VASA+ germ cells (D'). Inserts show enlargement of SIV RNA positive cells co-stained for cell markers. I: testicular interstitium; ST: seminiferous tubules. Scale bars = 20 µm. (G) SIV RNA+ cells were counted in a minimum of 30 tissue sections/experiment in 3 independent experiments on a primary-infected macaque MGT. Results show the mean positive cell number/tissue area +/− SEM.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2268241&req=5

pone-0001792-g004: SIV localization within the male genital tract.Detection of SIV positive cells in the seminal vesicles (A, C), testes (B–B', D–D'), epididymides (E, E') and prostate (F) of primary-infected macaques using immunohistochemistry for SIVp27 (A, B) and in situ hybridization (ISH) for SIV gag RNA (C–F). The phenotype of SIV positive cells was determined using ISH for SIV gag RNA (visualized as black silver grains) combined with immunostaining for cell markers (visualized as brown staining): combined ISH for viral RNA and immunostaining for CD3 revealed black silver grains clustered over brown cells in the seminal vesicles (C), prostate (F), and epididymis (E'), indicating infection of CD3+T lymphocytes. Co-labelling of SIV RNA+ cells with the myeloid cell marker CD68 was also observed, as shown here for the epididymis (E). In the testis, SIV RNA was detected within the interstitium in HLA-DR+ cells (D) and within the seminiferous tubules in VASA+ germ cells (D'). Inserts show enlargement of SIV RNA positive cells co-stained for cell markers. I: testicular interstitium; ST: seminiferous tubules. Scale bars = 20 µm. (G) SIV RNA+ cells were counted in a minimum of 30 tissue sections/experiment in 3 independent experiments on a primary-infected macaque MGT. Results show the mean positive cell number/tissue area +/− SEM.
Mentions: SIV p27 positive cells were found within the stroma and close to the secretory epithelium of the seminal vesicles (Figure 4A), prostate and epididymides (data not shown) of primary-infected animals. Positive staining was observed in the interstitium and seminiferous tubules of the testis (Figure 4B, B'). In situ hybridization for SIV gag RNA also revealed positive cells in the stroma and, occasionally, within the epithelium of the seminal vesicles (Figure 4C), epididymides (Figure 4E, E') and prostate (Figure 4F) of primary-infected animals. Within the testis, SIV RNA+cells were observed in the interstitium (Figure 4D), occasionally bordering the seminiferous tubules (data not shown). In all tissues, these infected cells were mainly T lymphocytes (60–97% of SIV RNA+ cells co-labeled with CD3, the highest proportion being found in the prostate and seminal vesicles) (Figure 4C, E', F), and some macrophages (0–25% of cells co-labeled with CD68, the highest rate being consistently found in the epididymis) (Figure 4E). Interestingly, in the testis SIV gag positive cells that never co-localized with either HLA-DR, CD3 or CD68 were occasionally observed within the seminiferous tubules (Figure 4D') (on average 1 positive cell for 300 seminiferous tubules). These positive cells systematically co-localized with VASA, a specific germ cell marker [42]. Their distribution and localization, from the base to the middle of the tubules, suggested an association of SIV with pre-meiotic and meiotic germ cells. In chronically infected macaques, the same pattern of SIV localization was observed, but fewer cells were affected. Importantly, however, a few SIV+ cells were detected within the MGT of an animal with undetectable blood viremia (data not shown). A quantitative measurement of the number of SIV+ cells per tissue surface in a primary-infected animal evidenced that seminal vesicles and prostate displayed a higher number of positive cells when compared to the epididymides and testes (Figure 4G).

Bottom Line: We demonstrate for the first time the presence of SIV in the testes, epididymides, prostate and seminal vesicles as early as 14 days post-inoculation.The prostate and seminal vesicles appear to be the most efficiently infected reproductive organs, followed by the epididymides and testes.Within the male genital tract, mostly T lymphocytes and a small number of germ cells harbour SIV antigens and RNA.

View Article: PubMed Central - PubMed

Affiliation: INSERM U625, Rennes, University of Rennes I, Groupe d'Etude de la Reproduction chez l'Homme et les Mammifères, IFR 140, Campus de Beaulieu, Rennes, France.

ABSTRACT

Background: Although indirect evidence suggests the male genital tract as a possible source of persistent HIV shedding in semen during antiretroviral therapy, this phenomenon is poorly understood due to the difficulty of sampling semen-producing organs in HIV+ asymptomatic individuals.

Methodology/principal findings: Using a range of molecular and cell biological techniques, this study investigates SIV infection within reproductive organs of macaques during the acute and chronic stages of the disease. We demonstrate for the first time the presence of SIV in the testes, epididymides, prostate and seminal vesicles as early as 14 days post-inoculation. This infection persists throughout the chronic stage and positively correlates with blood viremia. The prostate and seminal vesicles appear to be the most efficiently infected reproductive organs, followed by the epididymides and testes. Within the male genital tract, mostly T lymphocytes and a small number of germ cells harbour SIV antigens and RNA. In contrast to the other organs studied, the testis does not display an immune response to the infection. Testosteronemia is transiently increased during the early phase of the infection but spermatogenesis remains unaffected.

Conclusions/significance: The present study reveals that SIV infection of the macaque male genital tract is an early event and that semen-producing organs display differential infection levels and immune responses. These results help elucidate the origin of HIV in semen and constitute an essential base to improving the design of antiretroviral therapies to eradicate virus from semen.

Show MeSH
Related in: MedlinePlus