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Regulatory Cell Populations in Relapsing-Remitting Multiple Sclerosis (RRMS) Patients: Effect of Disease Activity and Treatment Regimens

View Article: PubMed Central - PubMed

ABSTRACT

Multiple sclerosis (MS) is a demyelinating disease of the central nervous system (CNS) of autoimmune etiology that results from an imbalance between CNS-specific T effector cells and peripheral suppressive mechanisms mediated by regulatory cells (RC). In this research, we collected blood samples from 83 relapsing remitting MS (RRMS) patients and 45 healthy persons (HC), to assess the sizes of their RC populations, including CD4+CD25highFoxp3+ (nTregs), CD3+CD4+HLA−G+, CD3+CD8+CD28−, CD3+CD56+, and CD56bright cells, and how RC are affected by disease activity (acute phase or remission) and types of treatment (methylprednisolone, interferon, or natalizumab). In addition, we isolated peripheral blood mononuclear cells (PBMC) and cultured them with peptides mapping to myelin antigens, to determine RC responsiveness to autoantigens. The results showed decreased levels of nTregs in patients in the acute phase ± methylprednisolone and in remission + natalizumab, but HC levels in patients in remission or receiving interferon. Patients + interferon had the highest levels of CD3+CD4+HLA−G+ and CD3+CD8+CD28− RC, and patients in the acute phase + methylprednisolone the lowest. Patients in remission had the highest levels of CD3+CD56+, and patients in remission + natalizumab the highest levels of CD56bright cells. Only nTregs responded to autoantigens in culture, regardless of disease activity or treatment. The highest suppressive activity was exhibited by nTregs from patients in remission. In conclusion, in RRMS disease activity and type of treatment affect different RC populations. nTregs respond to myelin antigens, indicating that it is possible to restore immunological tolerance through nTreg induction.

No MeSH data available.


Flow cytometric analysis to determine CD3+CD4+HLA−G+ RC levels in human peripheral blood. A representative analysis is shown for one healthy control (A1–D1) and one RRMS patient (A2–D2); The WBC were gated on lymphocytes, based on forward and side light scatter (A1,A2) and analyzed for CD3 (B1,B2), CD4 (C1,C2) and HLA-G expression (D1,D2); The numbers in the dot plots indicate the percentage of gated cells expressing the relevant marker. The tables underneath show the absolute number of cells in each population analyzed; Bottom graph: The results of the analysis of all patients (n = 83) and controls (HC, n = 45). For abbreviations see legend of Figure 1. * p = 0.05, ** p = 0.01, *** p = 0.006.
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ijms-17-01398-f002: Flow cytometric analysis to determine CD3+CD4+HLA−G+ RC levels in human peripheral blood. A representative analysis is shown for one healthy control (A1–D1) and one RRMS patient (A2–D2); The WBC were gated on lymphocytes, based on forward and side light scatter (A1,A2) and analyzed for CD3 (B1,B2), CD4 (C1,C2) and HLA-G expression (D1,D2); The numbers in the dot plots indicate the percentage of gated cells expressing the relevant marker. The tables underneath show the absolute number of cells in each population analyzed; Bottom graph: The results of the analysis of all patients (n = 83) and controls (HC, n = 45). For abbreviations see legend of Figure 1. * p = 0.05, ** p = 0.01, *** p = 0.006.

Mentions: The levels of the other regulatory cell populations studied in the peripheral blood of MS patients and HC, are shown in Figure 2, Figure 3 and Figure 4. The percentage of CD3+CD4+HLA−G+ T-cells was higher in AP-IFN patients compared to all other groups, with the difference reaching statistical significance between AP-IFN and HC, AP-MP, and Rem-NATA patients (Figure 2).


Regulatory Cell Populations in Relapsing-Remitting Multiple Sclerosis (RRMS) Patients: Effect of Disease Activity and Treatment Regimens
Flow cytometric analysis to determine CD3+CD4+HLA−G+ RC levels in human peripheral blood. A representative analysis is shown for one healthy control (A1–D1) and one RRMS patient (A2–D2); The WBC were gated on lymphocytes, based on forward and side light scatter (A1,A2) and analyzed for CD3 (B1,B2), CD4 (C1,C2) and HLA-G expression (D1,D2); The numbers in the dot plots indicate the percentage of gated cells expressing the relevant marker. The tables underneath show the absolute number of cells in each population analyzed; Bottom graph: The results of the analysis of all patients (n = 83) and controls (HC, n = 45). For abbreviations see legend of Figure 1. * p = 0.05, ** p = 0.01, *** p = 0.006.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037678&req=5

ijms-17-01398-f002: Flow cytometric analysis to determine CD3+CD4+HLA−G+ RC levels in human peripheral blood. A representative analysis is shown for one healthy control (A1–D1) and one RRMS patient (A2–D2); The WBC were gated on lymphocytes, based on forward and side light scatter (A1,A2) and analyzed for CD3 (B1,B2), CD4 (C1,C2) and HLA-G expression (D1,D2); The numbers in the dot plots indicate the percentage of gated cells expressing the relevant marker. The tables underneath show the absolute number of cells in each population analyzed; Bottom graph: The results of the analysis of all patients (n = 83) and controls (HC, n = 45). For abbreviations see legend of Figure 1. * p = 0.05, ** p = 0.01, *** p = 0.006.
Mentions: The levels of the other regulatory cell populations studied in the peripheral blood of MS patients and HC, are shown in Figure 2, Figure 3 and Figure 4. The percentage of CD3+CD4+HLA−G+ T-cells was higher in AP-IFN patients compared to all other groups, with the difference reaching statistical significance between AP-IFN and HC, AP-MP, and Rem-NATA patients (Figure 2).

View Article: PubMed Central - PubMed

ABSTRACT

Multiple sclerosis (MS) is a demyelinating disease of the central nervous system (CNS) of autoimmune etiology that results from an imbalance between CNS-specific T effector cells and peripheral suppressive mechanisms mediated by regulatory cells (RC). In this research, we collected blood samples from 83 relapsing remitting MS (RRMS) patients and 45 healthy persons (HC), to assess the sizes of their RC populations, including CD4+CD25highFoxp3+ (nTregs), CD3+CD4+HLA−G+, CD3+CD8+CD28−, CD3+CD56+, and CD56bright cells, and how RC are affected by disease activity (acute phase or remission) and types of treatment (methylprednisolone, interferon, or natalizumab). In addition, we isolated peripheral blood mononuclear cells (PBMC) and cultured them with peptides mapping to myelin antigens, to determine RC responsiveness to autoantigens. The results showed decreased levels of nTregs in patients in the acute phase ± methylprednisolone and in remission + natalizumab, but HC levels in patients in remission or receiving interferon. Patients + interferon had the highest levels of CD3+CD4+HLA−G+ and CD3+CD8+CD28− RC, and patients in the acute phase + methylprednisolone the lowest. Patients in remission had the highest levels of CD3+CD56+, and patients in remission + natalizumab the highest levels of CD56bright cells. Only nTregs responded to autoantigens in culture, regardless of disease activity or treatment. The highest suppressive activity was exhibited by nTregs from patients in remission. In conclusion, in RRMS disease activity and type of treatment affect different RC populations. nTregs respond to myelin antigens, indicating that it is possible to restore immunological tolerance through nTreg induction.

No MeSH data available.