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B cell follicle sanctuary permits persistent productive simian immunodeficiency virus infection in elite controllers.

Fukazawa Y, Lum R, Okoye AA, Park H, Matsuda K, Bae JY, Hagen SI, Shoemaker R, Deleage C, Lucero C, Morcock D, Swanson T, Legasse AW, Axthelm MK, Hesselgesser J, Geleziunas R, Hirsch VM, Edlefsen PT, Piatak M, Estes JD, Lifson JD, Picker LJ - Nat. Med. (2015)

Bottom Line: Chronic-phase HIV and simian immunodeficiency virus (SIV) replication is reduced by as much as 10,000-fold in elite controllers (ECs) compared with typical progressors (TPs), but sufficient viral replication persists in EC tissues to allow viral sequence evolution and induce excess immune activation.Here we show that productive SIV infection in rhesus monkey ECs, but not TPs, is markedly restricted to CD4(+) follicular helper T (TFH) cells, suggesting that these EC monkeys' highly effective SIV-specific CD8(+) T cells can effectively clear productive SIV infection from extrafollicular sites, but their relative exclusion from B cell follicles prevents their elimination of productively infected TFH cells.Thus, B cell follicles constitute 'sanctuaries' for persistent SIV replication in the presence of potent anti-viral CD8(+) T cell responses, potentially complicating efforts to cure HIV infection with therapeutic vaccination or T cell immunotherapy.

View Article: PubMed Central - PubMed

Affiliation: 1] Vaccine and Gene Therapy Institute, Oregon Health &Science University, Beaverton, Oregon, USA. [2] Oregon National Primate Research Center, Oregon Health &Science University, Beaverton, Oregon, USA.

ABSTRACT
Chronic-phase HIV and simian immunodeficiency virus (SIV) replication is reduced by as much as 10,000-fold in elite controllers (ECs) compared with typical progressors (TPs), but sufficient viral replication persists in EC tissues to allow viral sequence evolution and induce excess immune activation. Here we show that productive SIV infection in rhesus monkey ECs, but not TPs, is markedly restricted to CD4(+) follicular helper T (TFH) cells, suggesting that these EC monkeys' highly effective SIV-specific CD8(+) T cells can effectively clear productive SIV infection from extrafollicular sites, but their relative exclusion from B cell follicles prevents their elimination of productively infected TFH cells. CD8(+) lymphocyte depletion in EC monkeys resulted in a dramatic re-distribution of productive SIV infection to non-TFH cells, with restriction of productive infection to TFH cells resuming upon CD8(+) T cell recovery. Thus, B cell follicles constitute 'sanctuaries' for persistent SIV replication in the presence of potent anti-viral CD8(+) T cell responses, potentially complicating efforts to cure HIV infection with therapeutic vaccination or T cell immunotherapy.

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Levels of productive SIV infection within CD4+ memory T cell subsets differ in chronic phase attenuated vs. wildtype (WT) SIVmac239 infection(a) Plasma viral load profiles of representative SIVmac239 and SIVmac239Δnef infections. (b,c) Detection of replication-competent SIV within PD-1/CD200-defined, CD4+ memory T cell populations (TFH vs. non-TFH; 104 sorted cells, isolated as shown in Suppl. Fig. 1) obtained at the designated time points post-infection from Rh27617 (b; WT SIVmac239) and Rh25653 (c; SIVmac239Δnef) after 17 days of co-culture with CEMx174 cells (using flow cytometric analysis of intracellular SIV-Gag p27 to demonstrate CEMx174 cell infection; %Gag+ cell shown). See Suppl. Fig. 3 for similar analysis of Rh26310 (WT SIVmac239) and Rh25714 (SIVmac239Δnef).
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Figure 1: Levels of productive SIV infection within CD4+ memory T cell subsets differ in chronic phase attenuated vs. wildtype (WT) SIVmac239 infection(a) Plasma viral load profiles of representative SIVmac239 and SIVmac239Δnef infections. (b,c) Detection of replication-competent SIV within PD-1/CD200-defined, CD4+ memory T cell populations (TFH vs. non-TFH; 104 sorted cells, isolated as shown in Suppl. Fig. 1) obtained at the designated time points post-infection from Rh27617 (b; WT SIVmac239) and Rh25653 (c; SIVmac239Δnef) after 17 days of co-culture with CEMx174 cells (using flow cytometric analysis of intracellular SIV-Gag p27 to demonstrate CEMx174 cell infection; %Gag+ cell shown). See Suppl. Fig. 3 for similar analysis of Rh26310 (WT SIVmac239) and Rh25714 (SIVmac239Δnef).

Mentions: Although CD4+ TFH have been shown to constitute a major, and often disproportionate, component of productively HIV- and SIV-infected cell populations during chronic phase infection, it is unclear whether preferential viral targeting of this subset is based on enhanced susceptibility to infection, a longer intrinsic lifespan of infected cells, location in a particularly conducive environment for productive infection, the relative expansion of CD4+ TFH vs. non-TFH in progressive infection, or preferential shielding of productively infected CD4+ TFH from immune elimination29–33. To initially address this question, we quantified replication-competent virus by co-culture analysis of sorted CD4+ TFH vs. non-TFH memory T cells from lymph nodes (LNs) of four rhesus monkeys over the course of controlled LAV (nef-deleted SIVmac239) vs. progressive wildtype (WT) SIVmac239 infection. CD4+ TFH, delineated by high co-expression of PD1 and CD200 on CD4+, CD95high memory cells21,34, were compared to non-TFH CD4+ memory T cells that lacked PD1 and CD200 expression completely (PD-1neg/CD200neg) or expressed these markers at low level (PD-1dim/CD200dim; Suppl. Figs. 1 and 2). During acute LAV infection, and all phases of progressive WT SIV infection, replication-competent virus was present at similar levels in both CD4+ TFH and non-TFH memory T cells (Fig. 1a–c; Suppl. Fig. 3). However, after the onset of immunologic (primarily CD8+ T cell-mediated) control of LAV replication, replication-competent virus could only be isolated from TFH (Fig. 1c). These data suggest that the preferential localization of replication-competent LAV within CD4+ TFH noted in our previous report21 was related to the development of effective CD8+ T cell responses, which appear to preferentially restrict LAV replication in the T cell zone of the LN paracortex relative to the B cell follicles.


B cell follicle sanctuary permits persistent productive simian immunodeficiency virus infection in elite controllers.

Fukazawa Y, Lum R, Okoye AA, Park H, Matsuda K, Bae JY, Hagen SI, Shoemaker R, Deleage C, Lucero C, Morcock D, Swanson T, Legasse AW, Axthelm MK, Hesselgesser J, Geleziunas R, Hirsch VM, Edlefsen PT, Piatak M, Estes JD, Lifson JD, Picker LJ - Nat. Med. (2015)

Levels of productive SIV infection within CD4+ memory T cell subsets differ in chronic phase attenuated vs. wildtype (WT) SIVmac239 infection(a) Plasma viral load profiles of representative SIVmac239 and SIVmac239Δnef infections. (b,c) Detection of replication-competent SIV within PD-1/CD200-defined, CD4+ memory T cell populations (TFH vs. non-TFH; 104 sorted cells, isolated as shown in Suppl. Fig. 1) obtained at the designated time points post-infection from Rh27617 (b; WT SIVmac239) and Rh25653 (c; SIVmac239Δnef) after 17 days of co-culture with CEMx174 cells (using flow cytometric analysis of intracellular SIV-Gag p27 to demonstrate CEMx174 cell infection; %Gag+ cell shown). See Suppl. Fig. 3 for similar analysis of Rh26310 (WT SIVmac239) and Rh25714 (SIVmac239Δnef).
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Figure 1: Levels of productive SIV infection within CD4+ memory T cell subsets differ in chronic phase attenuated vs. wildtype (WT) SIVmac239 infection(a) Plasma viral load profiles of representative SIVmac239 and SIVmac239Δnef infections. (b,c) Detection of replication-competent SIV within PD-1/CD200-defined, CD4+ memory T cell populations (TFH vs. non-TFH; 104 sorted cells, isolated as shown in Suppl. Fig. 1) obtained at the designated time points post-infection from Rh27617 (b; WT SIVmac239) and Rh25653 (c; SIVmac239Δnef) after 17 days of co-culture with CEMx174 cells (using flow cytometric analysis of intracellular SIV-Gag p27 to demonstrate CEMx174 cell infection; %Gag+ cell shown). See Suppl. Fig. 3 for similar analysis of Rh26310 (WT SIVmac239) and Rh25714 (SIVmac239Δnef).
Mentions: Although CD4+ TFH have been shown to constitute a major, and often disproportionate, component of productively HIV- and SIV-infected cell populations during chronic phase infection, it is unclear whether preferential viral targeting of this subset is based on enhanced susceptibility to infection, a longer intrinsic lifespan of infected cells, location in a particularly conducive environment for productive infection, the relative expansion of CD4+ TFH vs. non-TFH in progressive infection, or preferential shielding of productively infected CD4+ TFH from immune elimination29–33. To initially address this question, we quantified replication-competent virus by co-culture analysis of sorted CD4+ TFH vs. non-TFH memory T cells from lymph nodes (LNs) of four rhesus monkeys over the course of controlled LAV (nef-deleted SIVmac239) vs. progressive wildtype (WT) SIVmac239 infection. CD4+ TFH, delineated by high co-expression of PD1 and CD200 on CD4+, CD95high memory cells21,34, were compared to non-TFH CD4+ memory T cells that lacked PD1 and CD200 expression completely (PD-1neg/CD200neg) or expressed these markers at low level (PD-1dim/CD200dim; Suppl. Figs. 1 and 2). During acute LAV infection, and all phases of progressive WT SIV infection, replication-competent virus was present at similar levels in both CD4+ TFH and non-TFH memory T cells (Fig. 1a–c; Suppl. Fig. 3). However, after the onset of immunologic (primarily CD8+ T cell-mediated) control of LAV replication, replication-competent virus could only be isolated from TFH (Fig. 1c). These data suggest that the preferential localization of replication-competent LAV within CD4+ TFH noted in our previous report21 was related to the development of effective CD8+ T cell responses, which appear to preferentially restrict LAV replication in the T cell zone of the LN paracortex relative to the B cell follicles.

Bottom Line: Chronic-phase HIV and simian immunodeficiency virus (SIV) replication is reduced by as much as 10,000-fold in elite controllers (ECs) compared with typical progressors (TPs), but sufficient viral replication persists in EC tissues to allow viral sequence evolution and induce excess immune activation.Here we show that productive SIV infection in rhesus monkey ECs, but not TPs, is markedly restricted to CD4(+) follicular helper T (TFH) cells, suggesting that these EC monkeys' highly effective SIV-specific CD8(+) T cells can effectively clear productive SIV infection from extrafollicular sites, but their relative exclusion from B cell follicles prevents their elimination of productively infected TFH cells.Thus, B cell follicles constitute 'sanctuaries' for persistent SIV replication in the presence of potent anti-viral CD8(+) T cell responses, potentially complicating efforts to cure HIV infection with therapeutic vaccination or T cell immunotherapy.

View Article: PubMed Central - PubMed

Affiliation: 1] Vaccine and Gene Therapy Institute, Oregon Health &Science University, Beaverton, Oregon, USA. [2] Oregon National Primate Research Center, Oregon Health &Science University, Beaverton, Oregon, USA.

ABSTRACT
Chronic-phase HIV and simian immunodeficiency virus (SIV) replication is reduced by as much as 10,000-fold in elite controllers (ECs) compared with typical progressors (TPs), but sufficient viral replication persists in EC tissues to allow viral sequence evolution and induce excess immune activation. Here we show that productive SIV infection in rhesus monkey ECs, but not TPs, is markedly restricted to CD4(+) follicular helper T (TFH) cells, suggesting that these EC monkeys' highly effective SIV-specific CD8(+) T cells can effectively clear productive SIV infection from extrafollicular sites, but their relative exclusion from B cell follicles prevents their elimination of productively infected TFH cells. CD8(+) lymphocyte depletion in EC monkeys resulted in a dramatic re-distribution of productive SIV infection to non-TFH cells, with restriction of productive infection to TFH cells resuming upon CD8(+) T cell recovery. Thus, B cell follicles constitute 'sanctuaries' for persistent SIV replication in the presence of potent anti-viral CD8(+) T cell responses, potentially complicating efforts to cure HIV infection with therapeutic vaccination or T cell immunotherapy.

Show MeSH
Related in: MedlinePlus