Differential expression of CD26 on virus-specific CD8(+) T cells during active, latent and resolved infection.

The hallmark of effective establishment of immune memory is the long-term memory cell that persists in the absence of antigen. To explore its characteristics, we investigated the differences between a resolved successful immune response, such as after influenza (flu) vaccination, and the state of chronic infection with persistent antigen, such as with cytomegalovirus (CMV), Epstein-Barr virus (EBV) or human immunodeficiency virus (HIV), which leads to defective T-cell memory. Immunophenotypic analyses using multi-parameter flow cytometry and tetramer technology identified a unique pattern of CD26(high) expression among influenza-specific CD8(+) T cells, but not among CD8(+) T cells specific for CMV, EBV (three different epitopes) or HIV. The median percentage of CD8(+) T cells expressing CD26 was 95.5% for influenza, but for cells specific for CMV, EBV and HIV it was 10.5%, 12%-19%, and 13.2%, respectively. These findings suggest that expression of CD26(high) may be a characteristic of a memory cell. CD26(high) expression correlates with expression of CD127, a marker of memory cells. Furthermore, CD26(high) cells can produce interleukin-2. These findings offer insight into the dynamics of T-cell differentiation, and they may offer a specific marker of a successfully developed memory CD8(+) T cell, that of CD26(high). This marker has the potential to be useful in studies of immune responses to infectious agents, and to new vaccine candidates.

Intravenous administration of human bone marrow stromal cells induces angiogenesis in the ischemic boundary zone after stroke in rats.

We tested the hypothesis that intravenous infusion of human bone marrow stromal cells (hMSCs) promotes vascular endothelial growth factor (VEGF) secretion, VEGF receptor 2 (VEGFR2) expression and angiogenesis in the ischemic boundary zone (IBZ) after stroke. hMSCs (1x10(6)) were intravenously injected into rats 24 hours after middle cerebral artery occlusion (MCAo). Laser scanning confocal microscopy (LSCM), immunohistochemistry and ELISA were performed to assay angiogenesis and levels of human and rat VEGF in the host brain, respectively. In addition, capillary-like tube formation was measured using mouse brain-derived endothelial cells (MBDECs). Morphological and three dimensional image analyses revealed significant (P<0.05) increases in numbers of enlarged and thin walled blood vessels and numbers of newly formed capillaries at the boundary of the ischemic lesion in rats (n=12) treated with hMSCs compared with numbers in rats (n=12) treated with PBS. ELISA measurements showed that treatment with hMSCs significantly (P<0.05) raised endogenous rat VEGF levels in the IBZ from 10.5+/-1.7 ng/mL in the control group to 17.5+/-1.6 ng/mL in the hMSC-treated group. In addition, treatment with hMSCs increased endogenous VEGFR2 immunoreactivity. In vitro, when MBDECs were incubated with the supernatant obtained from cultured hMSCs, capillary-like tube formation was significantly (P<0.01) induced. However, hMSC-induced capillary-like tube formation was significantly (P<0.01) inhibited when the endothelial cells were incubated with the supernatant from hMSCs in the presence of a neutralizing anti-VEGFR2. These data suggest that treatment of stroke with hMSCs enhances angiogenesis in the host brain and hMSC-enhanced angiogenesis is mediated by increases in levels of endogenous rat VEGF and VEGFR2.

Breast milk CD4+ T cells express high levels of C chemokine receptor 5 and CXC chemokine receptor 4 and are preserved in HIV-infected mothers receiving highly active antiretroviral therapy.

BACKGROUND: Transmission of human immunodeficiency virus (HIV) to the infant through breast-feeding is a major problem worldwide; however, the biological circumstances of such transmission remain unclear. Little characterization of breast milk CD4(+) T lymphocytes has been done so far. METHODS: We performed a detailed immunophenotypic analysis of T lymphocytes in the breast milk, compared with the blood, of HIV-uninfected (n=9) and HIV-infected (n=10) women receiving highly active antiretroviral therapy, by use of multiparameter flow cytometry. Descriptive statistics and nonparametric comparisons were performed using SAS software (version 9.1; SAS Institute). RESULTS: In uninfected women, 44%-78% of breast milk CD4(+) T cells expressed the C chemokine receptor 5 (CCR5), whereas 26%-73% of cells coexpressed CCR5 and CXC chemokine receptor 4 (CXCR4). In contrast, only 7%-20% of peripheral blood CD4(+) T cells expressed CCR5 and 1%-20% coexpressed CCR5 and CXCR4. The level of CCR5 expression in CD4(+) T cells in breast milk was higher than in blood. In HIV-infected women, the high frequency of CD4(+)CCR5(+) T cells in breast milk was preserved. CONCLUSIONS: A majority of CD4(+) T cells in breast milk express high levels of CCR5 and CXCR4. Unlike other mucosal immune sites, in which CD4(+)CCR5(+) T cells are rapidly eliminated by HIV, these cells are preserved in breast milk during HIV infection.

The AIDS resistance of naturally SIV-infected sooty mangabeys is independent of cellular immunity to the virus.

In contrast to human immunodeficiency virus (HIV)-infected humans, natural hosts for simian immunodeficiency virus (SIV) very rarely progress to acquired immunodeficiency syndrome (AIDS). While the mechanisms underlying this disease resistance are still poorly understood, a consistent feature of natural SIV infection is the absence of the generalized immune activation associated with HIV infection. To investigate the immunologic mechanisms underlying the absence of AIDS in SIV-infected sooty mangabeys (SMs), a natural host species, we performed a detailed analysis of the SIV-specific cellular immune responses in 110 SIV-infected SMs. We found that while SIV-specific T-cell responses are detectable in the majority of animals, their magnitude and breadth are, in fact, lower than what has been described in HIV-infected humans, both in terms of cytokine production (ie, IFN-gamma, TNF-alpha, and IL-2) and degranulation (ie, CD107a expression). Of importance, SIV-specific T-cell responses were similarly low when either SIVmac239-derived peptides or autologous SIVsmm peptides were used as stimuli. No correlation was found between SIV-specific T-cell responses and either viral load or CD4+ T-cell count, or between these responses and markers of T-cell activation and proliferation. These findings indicate that the absence of AIDS in naturally SIV-infected sooty mangabeys is independent of a strong cellular immune response to the virus.

Expression of killer cell lectin-like receptor G1 on antigen-specific human CD8+ T lymphocytes during active, latent, and resolved infection and its relation with CD57.

Killer cell lectin-like receptor G1 (KLRG1) is one of several inhibitory killer cell lectin-like receptors expressed by NK cells and T lymphocytes, mainly CD8(+) effector/memory cells that can secrete cytokines but have poor proliferative capacity. Using multiparameter flow cytometry, we studied KLRG1 expression on CD8(+) T cells specific for epitopes of CMV, EBV, influenza, and HIV. Over 92% of CD8(+) cells specific for CMV or EBV expressed KLRG1 during the latent stage of these chronic infections. CD8(+) T cell cells specific for HIV epitopes were mostly (72-89%) KLRG1(+), even though not quite at the level of predominance noted with CMV or EBV. Lower frequency of KLRG1 expression was observed among CD8(+) cells specific for influenza (40-73%), a resolved infection without a latent stage. We further observed that CD8(+) cells expressing CD57, a marker of replicative senescence, also expressed KLRG1; however, a population of CD57(-)KLRG1(+) cells was also identified. This population may represent a "memory" phenotype, because they also expressed CD27, CD28, CCR7, and CD127. In contrast, CD57(+)KLRG1(+) cells did not express CD27, CD28, and CCR7, and expressed CD127 at a much lower frequency, indicating that they represent effector cells that are truly terminally differentiated. The combination of KLRG1 and CD57 expression might thus aid in refining functional characterization of CD8(+) T cell subsets.