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, 7 (3), 515-8

Secretory Leukocyte Protease Inhibitor Inhibits Infection of Monocytes and Lymphocytes With Human Immunodeficiency Virus Type 1 but Does Not Interfere With Transcytosis of Cell-Associated Virus Across Tight Epithelial Barriers

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Secretory Leukocyte Protease Inhibitor Inhibits Infection of Monocytes and Lymphocytes With Human Immunodeficiency Virus Type 1 but Does Not Interfere With Transcytosis of Cell-Associated Virus Across Tight Epithelial Barriers

H Hocini et al. Clin Diagn Lab Immunol.

Abstract

In the present study, we demonstrate that recombinant human secretory leukocyte protease inhibitor (rhSLPI) inhibits infection of lymphocyte- and monocyte-derived tumor cell lines and peripheral blood lymphocytes with laboratory-adapted isolates and with the primary isolate, NDK, of free human immunodeficiency virus type 1 (HIV-1). In contrast, rhSLPI did not exhibit inhibitory activity toward transcytosis of cell-associated HIV-1 through a tight monolayer of endometrial epithelial cells. These observations indicate that the inhibitory effect of SLPI is restricted to free HIV-1 in corporal fluids.

Figures

FIG. 1
FIG. 1
Immunohistochemical localization of SLPI in the endocervix. A sample of cervix tissue was obtained from a 35-year-old HIV-seronegative woman. Serial sections were saturated by treatment with 3% bovine serum albumin in phosphate-buffered saline (PBS) and further incubated with rabbit IgG anti-human SLPI (3 μg/ml) in PBS–0.1% bovine serum albumin. The sections were washed with PBS and revealed with biotinylated mouse Ig to rabbit Fcγ. After the sections had been washed, streptavidin-biotin-phosphatase alkaline complex was added for 30 min, and the slides were incubated with a chromogenic solution containing fast red. After the slides had been washed, they were counterstained with hematoxylin. Lung tissues were used as a positive control (not shown). The negative control included normal rabbit serum IgG instead of the anti-SLPI antibody. The secretory epithelium and glandular structures of the endocervix, but not the exocervix (not shown), showed marked immunoreactivities for SLPI (in red). Magnification, ×200.
FIG. 2
FIG. 2
Effect of rhSLPI on HIV-1 infection of lymphocyte tumor cell line SupT1, promonocyte-derived tumor cell line U937, and PBLs. (A) Free laboratory-adapted HIV-1 subtype A was incubated with rhSLPI (20 μg) (open circles) or RPMI 1640 (solid circles) for 1 h prior to addition to SupT1 cells (solid line) and further incubation for 2 h. The cells were then washed and cultured for 10 days. Cell supernatants were removed at 3- to 4-day intervals, and p24 antigen was quantitated in supernatants. Free HIV-1LAI was incubated with rhSLPI (open circles) or RPMI 1640 (closed circles) prior to addition to U937 cells (dotted line), as described above in the case of SupT1 cells. (B) Free primary isolate HIV-1 NDK was incubated with rhSLPI (20 μg) (open circles) or RPMI 1640 (solid circles) for 1 h at 37°C before infection of PBLs, as described above. Data are expressed as the mean ± standard error from the same experiment performed in duplicate.
FIG. 3
FIG. 3
Lack of inhibitory activity of rhSLPI on transcytosis through a tight epithelial monolayer of SupT1 cell-associated HIV-1 subtype A, of U937 cell-associated HIVLAI, and of the PBL-associated primary HIV-1 NDK isolate. Transcytosis was expressed as the percentage of p24 antigen recovered in the basolateral chamber in the presence of rhSLPI compared with the amount of p24 antigen recovered in the presence of irrelevant IgG. HIV transcytosis was markedly inhibited by immunopurified human polyclonal anti-gp160 IgG antibodies (positive control). No inhibition of transcytosis was observed after preincubation of HIV-infected cells with HIV-seronegative IgG, RPMI 1640 alone, or rhSLPI. Data are expressed as the mean percentage ± standard error in the same experiment performed in duplicate. Only data obtained with SLPI (20 μg) are shown.

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