Limits...
Blocking of integrins inhibits HIV-1 infection of human cervical mucosa immune cells with free and complement-opsonized virions.

Tjomsland V, Ellegård R, Kjölhede P, Wodlin NB, Hinkula J, Lifson JD, Larsson M - Eur. J. Immunol. (2013)

Bottom Line: Blockage of the α4-, β7-, and β1-integrins significantly inhibited HIV-1 infection of both DCs and CD4(+) T cells.We found a greater impairment of HIV-1 infection in DCs for complement-opsonized virions compared with that of free virions when αM/β2- and α4-integrins were blocked.We show that blocking of integrins decreases the HIV-1 infection of both mucosal DCs and CD4(+) T cells emigrating from the cervical tissues.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Virology, Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden.

Show MeSH

Related in: MedlinePlus

Blockade of CD4 and CCR5 decreases the infection of emigrating cervical mucosa DCs and T cells. Cervical tissue biopsies were infected with HIV-1BaL, either as free virions (F-HIV) or complement opsonized virions (C-HIV) that had been mock treated or incubated with b12 mAb, for 2 h at 37°C.The cervical tissue biopsies were pretreated for 30 min at 37°C with TAK779 followed by infection with different forms of HIV-1BaL, either free (F-HIV) or complement opsonized (C-HIV), for 2 h at 37°C. The tissues were washed and transferred to six-well plates and cultured for 3–6 days in the absence or continuous presence of b12 mAb or TAK779. The emigrating cells were harvested and stained with anti-CD1a, and anti-HIV-1 mAbs for DCs and anti-CD3, anti-CD4, and anti-HIV-1 mAbs for CD4+ T cells and level of infection was assessed by flow cytometry for HIV-1 p24 expression. (A, B) The level of HIV-1 infection after mock treatment or treatment with b12 in the different (A) DCs (N = 4–6) or (B) T cells (N = 4–7) was normalized with mock-treated F-HIV set as 100%. (C, D) The level of HIV-1 infection after mock or TAK779 treatment in the different (C) DC or (D) T-cell experiments was normalized with F-HIV set as 100% (N = 3–4). Statistical significance was tested using a two-sided paired t-test. *p < 0.05, **p < 0.001, ***p < 0.0001. Data are shown as mean + SEM of the indicated number of samples, each assessed in its own experiment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4231223&req=5

fig03: Blockade of CD4 and CCR5 decreases the infection of emigrating cervical mucosa DCs and T cells. Cervical tissue biopsies were infected with HIV-1BaL, either as free virions (F-HIV) or complement opsonized virions (C-HIV) that had been mock treated or incubated with b12 mAb, for 2 h at 37°C.The cervical tissue biopsies were pretreated for 30 min at 37°C with TAK779 followed by infection with different forms of HIV-1BaL, either free (F-HIV) or complement opsonized (C-HIV), for 2 h at 37°C. The tissues were washed and transferred to six-well plates and cultured for 3–6 days in the absence or continuous presence of b12 mAb or TAK779. The emigrating cells were harvested and stained with anti-CD1a, and anti-HIV-1 mAbs for DCs and anti-CD3, anti-CD4, and anti-HIV-1 mAbs for CD4+ T cells and level of infection was assessed by flow cytometry for HIV-1 p24 expression. (A, B) The level of HIV-1 infection after mock treatment or treatment with b12 in the different (A) DCs (N = 4–6) or (B) T cells (N = 4–7) was normalized with mock-treated F-HIV set as 100%. (C, D) The level of HIV-1 infection after mock or TAK779 treatment in the different (C) DC or (D) T-cell experiments was normalized with F-HIV set as 100% (N = 3–4). Statistical significance was tested using a two-sided paired t-test. *p < 0.05, **p < 0.001, ***p < 0.0001. Data are shown as mean + SEM of the indicated number of samples, each assessed in its own experiment.

Mentions: We investigated the infection in DCs and CD4+ T cells after blocking CD4 with the mAb b12, a neutralizing antibody that specifically targets the CD4 binding site on gp120, and CCR5 with the inhibitor TAK779. We mostly used the b12 experiment to establish the level of inhibition reached in this model by a known blocker of HIV-1 infection 24. b12 significantly decreased the infection of migrating DCs to 45% of unblocked control for F-HIV (p = 0.019) and to 62% of control for C-HIV (p = 0.028) (Fig. 3A). When b12 was used, infection of CD4+ T cells emigrating from cervical explants was decreased by 85% (p < 0.0001) for F-HIV and by 74% (p < 0.0001) for C-HIV (Fig. 3B). The CCR5 inhibitor TAK799 decreased the HIV-1 infection of emigrating DCs by 41% (p = 0.037) for F-HIV and 66% for C-HIV (p = 0.06) and of CD4+ T cells by 48% (p = 0.02) for F-HIV and 68% (p = 0.01) for C-HIV (Fig. 3C and D).


Blocking of integrins inhibits HIV-1 infection of human cervical mucosa immune cells with free and complement-opsonized virions.

Tjomsland V, Ellegård R, Kjölhede P, Wodlin NB, Hinkula J, Lifson JD, Larsson M - Eur. J. Immunol. (2013)

Blockade of CD4 and CCR5 decreases the infection of emigrating cervical mucosa DCs and T cells. Cervical tissue biopsies were infected with HIV-1BaL, either as free virions (F-HIV) or complement opsonized virions (C-HIV) that had been mock treated or incubated with b12 mAb, for 2 h at 37°C.The cervical tissue biopsies were pretreated for 30 min at 37°C with TAK779 followed by infection with different forms of HIV-1BaL, either free (F-HIV) or complement opsonized (C-HIV), for 2 h at 37°C. The tissues were washed and transferred to six-well plates and cultured for 3–6 days in the absence or continuous presence of b12 mAb or TAK779. The emigrating cells were harvested and stained with anti-CD1a, and anti-HIV-1 mAbs for DCs and anti-CD3, anti-CD4, and anti-HIV-1 mAbs for CD4+ T cells and level of infection was assessed by flow cytometry for HIV-1 p24 expression. (A, B) The level of HIV-1 infection after mock treatment or treatment with b12 in the different (A) DCs (N = 4–6) or (B) T cells (N = 4–7) was normalized with mock-treated F-HIV set as 100%. (C, D) The level of HIV-1 infection after mock or TAK779 treatment in the different (C) DC or (D) T-cell experiments was normalized with F-HIV set as 100% (N = 3–4). Statistical significance was tested using a two-sided paired t-test. *p < 0.05, **p < 0.001, ***p < 0.0001. Data are shown as mean + SEM of the indicated number of samples, each assessed in its own experiment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4231223&req=5

fig03: Blockade of CD4 and CCR5 decreases the infection of emigrating cervical mucosa DCs and T cells. Cervical tissue biopsies were infected with HIV-1BaL, either as free virions (F-HIV) or complement opsonized virions (C-HIV) that had been mock treated or incubated with b12 mAb, for 2 h at 37°C.The cervical tissue biopsies were pretreated for 30 min at 37°C with TAK779 followed by infection with different forms of HIV-1BaL, either free (F-HIV) or complement opsonized (C-HIV), for 2 h at 37°C. The tissues were washed and transferred to six-well plates and cultured for 3–6 days in the absence or continuous presence of b12 mAb or TAK779. The emigrating cells were harvested and stained with anti-CD1a, and anti-HIV-1 mAbs for DCs and anti-CD3, anti-CD4, and anti-HIV-1 mAbs for CD4+ T cells and level of infection was assessed by flow cytometry for HIV-1 p24 expression. (A, B) The level of HIV-1 infection after mock treatment or treatment with b12 in the different (A) DCs (N = 4–6) or (B) T cells (N = 4–7) was normalized with mock-treated F-HIV set as 100%. (C, D) The level of HIV-1 infection after mock or TAK779 treatment in the different (C) DC or (D) T-cell experiments was normalized with F-HIV set as 100% (N = 3–4). Statistical significance was tested using a two-sided paired t-test. *p < 0.05, **p < 0.001, ***p < 0.0001. Data are shown as mean + SEM of the indicated number of samples, each assessed in its own experiment.
Mentions: We investigated the infection in DCs and CD4+ T cells after blocking CD4 with the mAb b12, a neutralizing antibody that specifically targets the CD4 binding site on gp120, and CCR5 with the inhibitor TAK779. We mostly used the b12 experiment to establish the level of inhibition reached in this model by a known blocker of HIV-1 infection 24. b12 significantly decreased the infection of migrating DCs to 45% of unblocked control for F-HIV (p = 0.019) and to 62% of control for C-HIV (p = 0.028) (Fig. 3A). When b12 was used, infection of CD4+ T cells emigrating from cervical explants was decreased by 85% (p < 0.0001) for F-HIV and by 74% (p < 0.0001) for C-HIV (Fig. 3B). The CCR5 inhibitor TAK799 decreased the HIV-1 infection of emigrating DCs by 41% (p = 0.037) for F-HIV and 66% for C-HIV (p = 0.06) and of CD4+ T cells by 48% (p = 0.02) for F-HIV and 68% (p = 0.01) for C-HIV (Fig. 3C and D).

Bottom Line: Blockage of the α4-, β7-, and β1-integrins significantly inhibited HIV-1 infection of both DCs and CD4(+) T cells.We found a greater impairment of HIV-1 infection in DCs for complement-opsonized virions compared with that of free virions when αM/β2- and α4-integrins were blocked.We show that blocking of integrins decreases the HIV-1 infection of both mucosal DCs and CD4(+) T cells emigrating from the cervical tissues.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Virology, Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden.

Show MeSH
Related in: MedlinePlus