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Effective elicitation of human effector CD8+ T Cells in HLA-B*51:01 transgenic humanized mice after infection with HIV-1.

Sato Y, Nagata S, Takiguchi M - PLoS ONE (2012)

Bottom Line: However, it is well known that human CD8(+) T cells cannot differentiate into effector cells in immunodeficient mice transplanted with only human CD34(+) hematopoietic stem cells (HSCs), because human T cells are not educated by HLA in the mouse thymus.There were no differences in the frequency of late effector memory and effector subsets (CD27(low)CD28(-)CD45RA(+/-)CCR7(-) and CD27(-)CD28(-)CD45RA(+/-)CCR7(-), respectively) among human CD8(+) T cells and in that of human CD8(+) T cells expressing CX3CR1 and/or CXCR1 between hNOK/B51Tg and hNOK mice.These results suggest that hNOK/B51Tg mice had CD8(+) T cells that were capable of differentiating into effector T cells after viral antigen stimulation and had a greater ability to elicit effector CD8(+) T cells than hNOK ones.

View Article: PubMed Central - PubMed

Affiliation: Center for AIDS Research, Kumamoto University, 2-2-1 Honjo, Kumamoto, Japan.

ABSTRACT
Humanized mice are expected to be useful as small animal models for in vivo studies on the pathogenesis of infectious diseases. However, it is well known that human CD8(+) T cells cannot differentiate into effector cells in immunodeficient mice transplanted with only human CD34(+) hematopoietic stem cells (HSCs), because human T cells are not educated by HLA in the mouse thymus. We here established HLA-B*51:01 transgenic humanized mice by transplanting human CD34(+) HSCs into HLA-B*51:01 transgenic NOD/SCID/Jak3(-/-) mice (hNOK/B51Tg mice) and investigated whether human effector CD8(+) T cells would be elicited in the mice or in those infected with HIV-1 NL4-3. There were no differences in the frequency of late effector memory and effector subsets (CD27(low)CD28(-)CD45RA(+/-)CCR7(-) and CD27(-)CD28(-)CD45RA(+/-)CCR7(-), respectively) among human CD8(+) T cells and in that of human CD8(+) T cells expressing CX3CR1 and/or CXCR1 between hNOK/B51Tg and hNOK mice. In contrast, the frequency of late effector memory and effector CD8(+) T cell subsets and of those expressing CX3CR1 and/or CXCR1 was significantly higher in HIV-1-infected hNOK/B51Tg mice than in uninfected ones, whereas there was no difference in that of these subsets between HIV-1-infected and uninfected hNOK mice. These results suggest that hNOK/B51Tg mice had CD8(+) T cells that were capable of differentiating into effector T cells after viral antigen stimulation and had a greater ability to elicit effector CD8(+) T cells than hNOK ones.

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Establishment of a hNOK/B51Tg mouse model for the analysis of HIV-1 infections.NOK/B51Tg mice were established by backcrossing NOK mice with HLA-B*51:01 transgenic NOD/SCID mice, and then were analyzed for the expression of HLA-B51 molecules on their splenocytes. (A) Representative data on the expression of HLA-B51 molecules on splenocytes from a NOK/B51Tg mouse. Splenocytes were stained with TU109 mAB (black line) or isotype control (gray line). hNOK/B51Tg mice were analyzed for the frequency of reconstituted human cells at 10 weeks after the transplantation with human CD34+ cells. (B) Representative data and summary of the frequency of human CD19+ and CD3+ cells among the CD45+-gated subset in PBMCs (n = 18). (C) Representative data and summary of the frequency of human CD4+ and CD8+ T cells among CD45+/CD3+-gated subsets in PBMCs from hNOK/B51Tg mice (n = 18). hNOK/B51Tg mice were infected with HIV-1 at 14 weeks after the transplantation of CD34+ HSCs. (D) HIV-1-infected hNOK/B51Tg mice were analyzed for HIV-1 RNA and plasma viral load at 0, 2, 4, and 6 weeks post-infection (n = 11). (E) Representative data on human CD4+ and CD8+ T cell populations among CD45+/CD3+-gated subsets in PBMCs from an HIV-1-infected hNOK/B51Tg mouse (upper data) and from an uninfected one (lower data). (F) Summarized results on human CD4/CD8 T cell ratio at 0, 2, 4, and 6 weeks post-infection for PBMC from HIV-1-infected hNOK/B51Tg mice (n = 11, black circles) and from uninfected ones (n = 8, white circles). In uninfected hNOK/B51Tg mice, the proportion of human T cells in PBMC from the mice was observed from 14 weeks to 20 weeks after the transplantation. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected hNOK/B51Tg mice vs. uninfected ones). Error bars represent SEMs. (G) The number of human CD4+ T cells in peripheral blood from HIV-1-infecetd hNOK/B51Tg (n = 8, right data) and uninfected ones (n = 6, left data) was determined. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected hNOK/B51Tg mice at 2, 4, or 6 post-infection vs. hNOK/B51Tg mice before an HIV-1 infection).
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pone-0042776-g001: Establishment of a hNOK/B51Tg mouse model for the analysis of HIV-1 infections.NOK/B51Tg mice were established by backcrossing NOK mice with HLA-B*51:01 transgenic NOD/SCID mice, and then were analyzed for the expression of HLA-B51 molecules on their splenocytes. (A) Representative data on the expression of HLA-B51 molecules on splenocytes from a NOK/B51Tg mouse. Splenocytes were stained with TU109 mAB (black line) or isotype control (gray line). hNOK/B51Tg mice were analyzed for the frequency of reconstituted human cells at 10 weeks after the transplantation with human CD34+ cells. (B) Representative data and summary of the frequency of human CD19+ and CD3+ cells among the CD45+-gated subset in PBMCs (n = 18). (C) Representative data and summary of the frequency of human CD4+ and CD8+ T cells among CD45+/CD3+-gated subsets in PBMCs from hNOK/B51Tg mice (n = 18). hNOK/B51Tg mice were infected with HIV-1 at 14 weeks after the transplantation of CD34+ HSCs. (D) HIV-1-infected hNOK/B51Tg mice were analyzed for HIV-1 RNA and plasma viral load at 0, 2, 4, and 6 weeks post-infection (n = 11). (E) Representative data on human CD4+ and CD8+ T cell populations among CD45+/CD3+-gated subsets in PBMCs from an HIV-1-infected hNOK/B51Tg mouse (upper data) and from an uninfected one (lower data). (F) Summarized results on human CD4/CD8 T cell ratio at 0, 2, 4, and 6 weeks post-infection for PBMC from HIV-1-infected hNOK/B51Tg mice (n = 11, black circles) and from uninfected ones (n = 8, white circles). In uninfected hNOK/B51Tg mice, the proportion of human T cells in PBMC from the mice was observed from 14 weeks to 20 weeks after the transplantation. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected hNOK/B51Tg mice vs. uninfected ones). Error bars represent SEMs. (G) The number of human CD4+ T cells in peripheral blood from HIV-1-infecetd hNOK/B51Tg (n = 8, right data) and uninfected ones (n = 6, left data) was determined. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected hNOK/B51Tg mice at 2, 4, or 6 post-infection vs. hNOK/B51Tg mice before an HIV-1 infection).

Mentions: We established NOK/B51Tg mice by backcrossing NOK mice with HLA-B*51:01 transgenic NOD/SCID mice. First, the expression of HLA-B51 molecules on their splenocytes (Figure 1A) and PBMCs (data not shown) was confirmed. Next, we generated hNOK/B51Tg and hNOK mice by transplanting human CD34+ HSCs derived from 17 cord blood samples into the liver of newborn NOK/B51Tg and NOK mice, and then analyzed them for the population of reconstituted human cells among the PBMCs of the mice at 10 weeks after the transplantation (Table 1). Human CD45+ cells were detected in both hNOK/B51Tg and hNOK mice, and the frequency of these cells was not significantly different between the 2 groups of mice (Figure S1A). Human CD3+ T cells and CD19+ B cells were detected among the human CD45+-gated subset of PBMCs from hNOK/B51Tg mice (Figure 1B). The human CD3+ T cell population included both CD4+ and CD8+ T cells (Figure 1C). The frequency of their cells was not significantly different between hNOK/B51Tg and hNOK mice (Figure S1B–E). The human T cells were maintained in the mice at least for 20 weeks after the transplantation, and transgenic expression of HLA-B51 did not influence the proportion of human T cells in hNOK/B51Tg mice compared with that in hNOK mice (Figure 1E, uninfected hNOK/B51Tg mouse, Figure S2A, uninfected hNOK mouse). These results indicate that human T cells were generated and maintained in hNOK/B51Tg and hNOK mice. On the other hand, human CD14+ cells including dendritic cells (DCs) and macrophages were hardly detected in either mouse strain (data not shown).


Effective elicitation of human effector CD8+ T Cells in HLA-B*51:01 transgenic humanized mice after infection with HIV-1.

Sato Y, Nagata S, Takiguchi M - PLoS ONE (2012)

Establishment of a hNOK/B51Tg mouse model for the analysis of HIV-1 infections.NOK/B51Tg mice were established by backcrossing NOK mice with HLA-B*51:01 transgenic NOD/SCID mice, and then were analyzed for the expression of HLA-B51 molecules on their splenocytes. (A) Representative data on the expression of HLA-B51 molecules on splenocytes from a NOK/B51Tg mouse. Splenocytes were stained with TU109 mAB (black line) or isotype control (gray line). hNOK/B51Tg mice were analyzed for the frequency of reconstituted human cells at 10 weeks after the transplantation with human CD34+ cells. (B) Representative data and summary of the frequency of human CD19+ and CD3+ cells among the CD45+-gated subset in PBMCs (n = 18). (C) Representative data and summary of the frequency of human CD4+ and CD8+ T cells among CD45+/CD3+-gated subsets in PBMCs from hNOK/B51Tg mice (n = 18). hNOK/B51Tg mice were infected with HIV-1 at 14 weeks after the transplantation of CD34+ HSCs. (D) HIV-1-infected hNOK/B51Tg mice were analyzed for HIV-1 RNA and plasma viral load at 0, 2, 4, and 6 weeks post-infection (n = 11). (E) Representative data on human CD4+ and CD8+ T cell populations among CD45+/CD3+-gated subsets in PBMCs from an HIV-1-infected hNOK/B51Tg mouse (upper data) and from an uninfected one (lower data). (F) Summarized results on human CD4/CD8 T cell ratio at 0, 2, 4, and 6 weeks post-infection for PBMC from HIV-1-infected hNOK/B51Tg mice (n = 11, black circles) and from uninfected ones (n = 8, white circles). In uninfected hNOK/B51Tg mice, the proportion of human T cells in PBMC from the mice was observed from 14 weeks to 20 weeks after the transplantation. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected hNOK/B51Tg mice vs. uninfected ones). Error bars represent SEMs. (G) The number of human CD4+ T cells in peripheral blood from HIV-1-infecetd hNOK/B51Tg (n = 8, right data) and uninfected ones (n = 6, left data) was determined. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected hNOK/B51Tg mice at 2, 4, or 6 post-infection vs. hNOK/B51Tg mice before an HIV-1 infection).
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pone-0042776-g001: Establishment of a hNOK/B51Tg mouse model for the analysis of HIV-1 infections.NOK/B51Tg mice were established by backcrossing NOK mice with HLA-B*51:01 transgenic NOD/SCID mice, and then were analyzed for the expression of HLA-B51 molecules on their splenocytes. (A) Representative data on the expression of HLA-B51 molecules on splenocytes from a NOK/B51Tg mouse. Splenocytes were stained with TU109 mAB (black line) or isotype control (gray line). hNOK/B51Tg mice were analyzed for the frequency of reconstituted human cells at 10 weeks after the transplantation with human CD34+ cells. (B) Representative data and summary of the frequency of human CD19+ and CD3+ cells among the CD45+-gated subset in PBMCs (n = 18). (C) Representative data and summary of the frequency of human CD4+ and CD8+ T cells among CD45+/CD3+-gated subsets in PBMCs from hNOK/B51Tg mice (n = 18). hNOK/B51Tg mice were infected with HIV-1 at 14 weeks after the transplantation of CD34+ HSCs. (D) HIV-1-infected hNOK/B51Tg mice were analyzed for HIV-1 RNA and plasma viral load at 0, 2, 4, and 6 weeks post-infection (n = 11). (E) Representative data on human CD4+ and CD8+ T cell populations among CD45+/CD3+-gated subsets in PBMCs from an HIV-1-infected hNOK/B51Tg mouse (upper data) and from an uninfected one (lower data). (F) Summarized results on human CD4/CD8 T cell ratio at 0, 2, 4, and 6 weeks post-infection for PBMC from HIV-1-infected hNOK/B51Tg mice (n = 11, black circles) and from uninfected ones (n = 8, white circles). In uninfected hNOK/B51Tg mice, the proportion of human T cells in PBMC from the mice was observed from 14 weeks to 20 weeks after the transplantation. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected hNOK/B51Tg mice vs. uninfected ones). Error bars represent SEMs. (G) The number of human CD4+ T cells in peripheral blood from HIV-1-infecetd hNOK/B51Tg (n = 8, right data) and uninfected ones (n = 6, left data) was determined. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected hNOK/B51Tg mice at 2, 4, or 6 post-infection vs. hNOK/B51Tg mice before an HIV-1 infection).
Mentions: We established NOK/B51Tg mice by backcrossing NOK mice with HLA-B*51:01 transgenic NOD/SCID mice. First, the expression of HLA-B51 molecules on their splenocytes (Figure 1A) and PBMCs (data not shown) was confirmed. Next, we generated hNOK/B51Tg and hNOK mice by transplanting human CD34+ HSCs derived from 17 cord blood samples into the liver of newborn NOK/B51Tg and NOK mice, and then analyzed them for the population of reconstituted human cells among the PBMCs of the mice at 10 weeks after the transplantation (Table 1). Human CD45+ cells were detected in both hNOK/B51Tg and hNOK mice, and the frequency of these cells was not significantly different between the 2 groups of mice (Figure S1A). Human CD3+ T cells and CD19+ B cells were detected among the human CD45+-gated subset of PBMCs from hNOK/B51Tg mice (Figure 1B). The human CD3+ T cell population included both CD4+ and CD8+ T cells (Figure 1C). The frequency of their cells was not significantly different between hNOK/B51Tg and hNOK mice (Figure S1B–E). The human T cells were maintained in the mice at least for 20 weeks after the transplantation, and transgenic expression of HLA-B51 did not influence the proportion of human T cells in hNOK/B51Tg mice compared with that in hNOK mice (Figure 1E, uninfected hNOK/B51Tg mouse, Figure S2A, uninfected hNOK mouse). These results indicate that human T cells were generated and maintained in hNOK/B51Tg and hNOK mice. On the other hand, human CD14+ cells including dendritic cells (DCs) and macrophages were hardly detected in either mouse strain (data not shown).

Bottom Line: However, it is well known that human CD8(+) T cells cannot differentiate into effector cells in immunodeficient mice transplanted with only human CD34(+) hematopoietic stem cells (HSCs), because human T cells are not educated by HLA in the mouse thymus.There were no differences in the frequency of late effector memory and effector subsets (CD27(low)CD28(-)CD45RA(+/-)CCR7(-) and CD27(-)CD28(-)CD45RA(+/-)CCR7(-), respectively) among human CD8(+) T cells and in that of human CD8(+) T cells expressing CX3CR1 and/or CXCR1 between hNOK/B51Tg and hNOK mice.These results suggest that hNOK/B51Tg mice had CD8(+) T cells that were capable of differentiating into effector T cells after viral antigen stimulation and had a greater ability to elicit effector CD8(+) T cells than hNOK ones.

View Article: PubMed Central - PubMed

Affiliation: Center for AIDS Research, Kumamoto University, 2-2-1 Honjo, Kumamoto, Japan.

ABSTRACT
Humanized mice are expected to be useful as small animal models for in vivo studies on the pathogenesis of infectious diseases. However, it is well known that human CD8(+) T cells cannot differentiate into effector cells in immunodeficient mice transplanted with only human CD34(+) hematopoietic stem cells (HSCs), because human T cells are not educated by HLA in the mouse thymus. We here established HLA-B*51:01 transgenic humanized mice by transplanting human CD34(+) HSCs into HLA-B*51:01 transgenic NOD/SCID/Jak3(-/-) mice (hNOK/B51Tg mice) and investigated whether human effector CD8(+) T cells would be elicited in the mice or in those infected with HIV-1 NL4-3. There were no differences in the frequency of late effector memory and effector subsets (CD27(low)CD28(-)CD45RA(+/-)CCR7(-) and CD27(-)CD28(-)CD45RA(+/-)CCR7(-), respectively) among human CD8(+) T cells and in that of human CD8(+) T cells expressing CX3CR1 and/or CXCR1 between hNOK/B51Tg and hNOK mice. In contrast, the frequency of late effector memory and effector CD8(+) T cell subsets and of those expressing CX3CR1 and/or CXCR1 was significantly higher in HIV-1-infected hNOK/B51Tg mice than in uninfected ones, whereas there was no difference in that of these subsets between HIV-1-infected and uninfected hNOK mice. These results suggest that hNOK/B51Tg mice had CD8(+) T cells that were capable of differentiating into effector T cells after viral antigen stimulation and had a greater ability to elicit effector CD8(+) T cells than hNOK ones.

Show MeSH
Related in: MedlinePlus