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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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Phenotypic analysis of human CD4+ T cells in HIV-1-infected hNOK/B51Tg and hNOK mice as well as in uninfected ones.hNOK/B51Tg and hNOK mice were infected or not with HIV-1 at 14 weeks after the transplantation of CD34+ HSCs. Then human CD4+ T cells among PBMCs from HIV-1-infected and uninfected hNOK/B51Tg mice as well as HIV-1-infected and uninfected hNOK mice were analyzed for their expression of the following cell-surface markers: CD4, CD27, CD28, CD45RA, and CCR7. (A) Representative results of 5-color flow cytometric analysis of the human CD4+ T cell population in PBMCs from HIV-1-infected hNOK/B51Tg and HIV-1-infected hNOK mice at 6 weeks post-infection. (B) Same analysis as in “A” for uninfected hNOK/B51Tg and uninfected hNOK mice at 20 weeks after the transplantation of CD34+ HSCs. (C) Summarized results for the frequency of naive (CD27+CD28+CD45RA+CCR7+), central memory (CM:CD27+CD28+CD45RA−CCR7+), Th0 effector memory (Th0 EM:CD27+CD28+CD45RA−CCR7−), and Th1/2 effector memory (Th1/2 EM:CD27−CD28+CD45RA−CCR7−) plus effector (CD27−CD28−CD45RA−CCR7−) subsets among human CD4+ T cells in PBMCs from HIV-1-infected hNOK/B51Tg mice at 0, 2, 4, and 6 weeks post-infection (n = 11, black circles) and from uninfected ones at 14, 16, 18, and 20 weeks after the transplantation of CD34+ HSCs (n = 8, white circles) as well as those from HIV-1-infected hNOK mice (n = 8, black triangles) and from uninfected ones (n = 8, white triangles). Each symbol represents an individual mouse; and the mean value is shown as a horizontal solid line. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected mice vs. uninfected ones).
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pone-0042776-g002: Phenotypic analysis of human CD4+ T cells in HIV-1-infected hNOK/B51Tg and hNOK mice as well as in uninfected ones.hNOK/B51Tg and hNOK mice were infected or not with HIV-1 at 14 weeks after the transplantation of CD34+ HSCs. Then human CD4+ T cells among PBMCs from HIV-1-infected and uninfected hNOK/B51Tg mice as well as HIV-1-infected and uninfected hNOK mice were analyzed for their expression of the following cell-surface markers: CD4, CD27, CD28, CD45RA, and CCR7. (A) Representative results of 5-color flow cytometric analysis of the human CD4+ T cell population in PBMCs from HIV-1-infected hNOK/B51Tg and HIV-1-infected hNOK mice at 6 weeks post-infection. (B) Same analysis as in “A” for uninfected hNOK/B51Tg and uninfected hNOK mice at 20 weeks after the transplantation of CD34+ HSCs. (C) Summarized results for the frequency of naive (CD27+CD28+CD45RA+CCR7+), central memory (CM:CD27+CD28+CD45RA−CCR7+), Th0 effector memory (Th0 EM:CD27+CD28+CD45RA−CCR7−), and Th1/2 effector memory (Th1/2 EM:CD27−CD28+CD45RA−CCR7−) plus effector (CD27−CD28−CD45RA−CCR7−) subsets among human CD4+ T cells in PBMCs from HIV-1-infected hNOK/B51Tg mice at 0, 2, 4, and 6 weeks post-infection (n = 11, black circles) and from uninfected ones at 14, 16, 18, and 20 weeks after the transplantation of CD34+ HSCs (n = 8, white circles) as well as those from HIV-1-infected hNOK mice (n = 8, black triangles) and from uninfected ones (n = 8, white triangles). Each symbol represents an individual mouse; and the mean value is shown as a horizontal solid line. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected mice vs. uninfected ones).

Mentions: Human peripheral CD4+ T cells are classified into the following 5 major populations based on their expression of 4 cell-surface marker: CD27+CD28+CD45RA+CCR7+ (naive subset), CD27+CD28+CD45RA−CCR7+ (central memory subset: CM), CD27+CD28+CD45RA−CCR7− (Th0 effector memory subset: Th0 EM), CD27−CD28+CD45RA−CCR7− (Th1/2 effector memory subset: Th1/2 EM), and CD27−CD28−CD45RA−CCR7− (effector subset) [31]. To investigate the effect of an HIV-1 infection on the population of human CD4+ T cells in HIV-1-infected hNOK/B51Tg and hNOK mice, we analyzed the phenotype of these cells among PBMCs from the mice every 2 weeks post-infection. Representative data from HIV-1-infected hNOK/B51Tg and HIV-1-infected hNOK mice at 6 weeks post-infection (Figure 2A) and that from each uninfected ones at 20 weeks after the transplantation of CD34+ HSCs (Figure 2B), respectively, are shown; and summaries of the results from 11 HIV-1-infected and 8 uninfected hNOK/B51Tg mice as well as those from 8 HIV-1-infected and 8 uninfected hNOK mice are given in Figure 2C. The frequency of the CM subset among human CD4+ T cells from uninfected hNOK/B51Tg mice gradually increased every 2 weeks, whereas that for the HIV-1-infected hNOK/B51Tg mice was not altered during the HIV-1 infection. In addition, the frequency of Th1/2 EM and effector subsets among human CD4+ T cells for the infected mice gradually increased after the HIV-1 infection and was significantly higher at 6 weeks post-infection than that for the uninfected ones. These results show that the acute infection with the X4-tropic HIV-1 caused a depletion of the CM subset and a relative increase in the Th1/2 EM and effector subsets among human CD4+ T cells in the HIV-1-infected hNOK/B51Tg mice. On the other hand, there was no significant difference in the frequency of each subset among human CD4+ T cells between HIV-1-infected and uninfected hNOK mice (Figure 2C).


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)

Phenotypic analysis of human CD4+ T cells in HIV-1-infected hNOK/B51Tg and hNOK mice as well as in uninfected ones.hNOK/B51Tg and hNOK mice were infected or not with HIV-1 at 14 weeks after the transplantation of CD34+ HSCs. Then human CD4+ T cells among PBMCs from HIV-1-infected and uninfected hNOK/B51Tg mice as well as HIV-1-infected and uninfected hNOK mice were analyzed for their expression of the following cell-surface markers: CD4, CD27, CD28, CD45RA, and CCR7. (A) Representative results of 5-color flow cytometric analysis of the human CD4+ T cell population in PBMCs from HIV-1-infected hNOK/B51Tg and HIV-1-infected hNOK mice at 6 weeks post-infection. (B) Same analysis as in “A” for uninfected hNOK/B51Tg and uninfected hNOK mice at 20 weeks after the transplantation of CD34+ HSCs. (C) Summarized results for the frequency of naive (CD27+CD28+CD45RA+CCR7+), central memory (CM:CD27+CD28+CD45RA−CCR7+), Th0 effector memory (Th0 EM:CD27+CD28+CD45RA−CCR7−), and Th1/2 effector memory (Th1/2 EM:CD27−CD28+CD45RA−CCR7−) plus effector (CD27−CD28−CD45RA−CCR7−) subsets among human CD4+ T cells in PBMCs from HIV-1-infected hNOK/B51Tg mice at 0, 2, 4, and 6 weeks post-infection (n = 11, black circles) and from uninfected ones at 14, 16, 18, and 20 weeks after the transplantation of CD34+ HSCs (n = 8, white circles) as well as those from HIV-1-infected hNOK mice (n = 8, black triangles) and from uninfected ones (n = 8, white triangles). Each symbol represents an individual mouse; and the mean value is shown as a horizontal solid line. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected mice vs. uninfected ones).
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Related In: Results  -  Collection

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pone-0042776-g002: Phenotypic analysis of human CD4+ T cells in HIV-1-infected hNOK/B51Tg and hNOK mice as well as in uninfected ones.hNOK/B51Tg and hNOK mice were infected or not with HIV-1 at 14 weeks after the transplantation of CD34+ HSCs. Then human CD4+ T cells among PBMCs from HIV-1-infected and uninfected hNOK/B51Tg mice as well as HIV-1-infected and uninfected hNOK mice were analyzed for their expression of the following cell-surface markers: CD4, CD27, CD28, CD45RA, and CCR7. (A) Representative results of 5-color flow cytometric analysis of the human CD4+ T cell population in PBMCs from HIV-1-infected hNOK/B51Tg and HIV-1-infected hNOK mice at 6 weeks post-infection. (B) Same analysis as in “A” for uninfected hNOK/B51Tg and uninfected hNOK mice at 20 weeks after the transplantation of CD34+ HSCs. (C) Summarized results for the frequency of naive (CD27+CD28+CD45RA+CCR7+), central memory (CM:CD27+CD28+CD45RA−CCR7+), Th0 effector memory (Th0 EM:CD27+CD28+CD45RA−CCR7−), and Th1/2 effector memory (Th1/2 EM:CD27−CD28+CD45RA−CCR7−) plus effector (CD27−CD28−CD45RA−CCR7−) subsets among human CD4+ T cells in PBMCs from HIV-1-infected hNOK/B51Tg mice at 0, 2, 4, and 6 weeks post-infection (n = 11, black circles) and from uninfected ones at 14, 16, 18, and 20 weeks after the transplantation of CD34+ HSCs (n = 8, white circles) as well as those from HIV-1-infected hNOK mice (n = 8, black triangles) and from uninfected ones (n = 8, white triangles). Each symbol represents an individual mouse; and the mean value is shown as a horizontal solid line. Asterisks indicate statistically significant differences (*p<0.05, HIV-1-infected mice vs. uninfected ones).
Mentions: Human peripheral CD4+ T cells are classified into the following 5 major populations based on their expression of 4 cell-surface marker: CD27+CD28+CD45RA+CCR7+ (naive subset), CD27+CD28+CD45RA−CCR7+ (central memory subset: CM), CD27+CD28+CD45RA−CCR7− (Th0 effector memory subset: Th0 EM), CD27−CD28+CD45RA−CCR7− (Th1/2 effector memory subset: Th1/2 EM), and CD27−CD28−CD45RA−CCR7− (effector subset) [31]. To investigate the effect of an HIV-1 infection on the population of human CD4+ T cells in HIV-1-infected hNOK/B51Tg and hNOK mice, we analyzed the phenotype of these cells among PBMCs from the mice every 2 weeks post-infection. Representative data from HIV-1-infected hNOK/B51Tg and HIV-1-infected hNOK mice at 6 weeks post-infection (Figure 2A) and that from each uninfected ones at 20 weeks after the transplantation of CD34+ HSCs (Figure 2B), respectively, are shown; and summaries of the results from 11 HIV-1-infected and 8 uninfected hNOK/B51Tg mice as well as those from 8 HIV-1-infected and 8 uninfected hNOK mice are given in Figure 2C. The frequency of the CM subset among human CD4+ T cells from uninfected hNOK/B51Tg mice gradually increased every 2 weeks, whereas that for the HIV-1-infected hNOK/B51Tg mice was not altered during the HIV-1 infection. In addition, the frequency of Th1/2 EM and effector subsets among human CD4+ T cells for the infected mice gradually increased after the HIV-1 infection and was significantly higher at 6 weeks post-infection than that for the uninfected ones. These results show that the acute infection with the X4-tropic HIV-1 caused a depletion of the CM subset and a relative increase in the Th1/2 EM and effector subsets among human CD4+ T cells in the HIV-1-infected hNOK/B51Tg mice. On the other hand, there was no significant difference in the frequency of each subset among human CD4+ T cells between HIV-1-infected and uninfected hNOK mice (Figure 2C).

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