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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.


Related in: MedlinePlus

Flow cytometric analysis to determine CD4+CD25+Foxp3+ RC levels in human peripheral blood. A representative analysis is shown for one healthy control (A1–C1) and one relapsing remitting MS (RRMS) patient (A2–C2). The white blood cells (WBC) were gated on lymphocytes, based on forward and side light scatter (A1,A2) and analyzed for CD4 and CD25 expression (B1,B2); The double positive cells were analyzed further for Foxp3 expression (C1,C2); 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). AP-noRx (n = 13), patients in the acute phase of the disease without treatment; AP-MP (n = 17), patients in the acute phase under treatment with methylprednisolone; AP-IFN (n = 12), patients in the acute phase under treatment with interferon β; Rem-noRx (n = 15), patients in remission receiving no treatment; Rem-NATA (n = 26), patients in remission under treatment with natalizumab. * p = 0.04, ** p = 0.01, *** p = 0.002.
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ijms-17-01398-f001: Flow cytometric analysis to determine CD4+CD25+Foxp3+ RC levels in human peripheral blood. A representative analysis is shown for one healthy control (A1–C1) and one relapsing remitting MS (RRMS) patient (A2–C2). The white blood cells (WBC) were gated on lymphocytes, based on forward and side light scatter (A1,A2) and analyzed for CD4 and CD25 expression (B1,B2); The double positive cells were analyzed further for Foxp3 expression (C1,C2); 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). AP-noRx (n = 13), patients in the acute phase of the disease without treatment; AP-MP (n = 17), patients in the acute phase under treatment with methylprednisolone; AP-IFN (n = 12), patients in the acute phase under treatment with interferon β; Rem-noRx (n = 15), patients in remission receiving no treatment; Rem-NATA (n = 26), patients in remission under treatment with natalizumab. * p = 0.04, ** p = 0.01, *** p = 0.002.

Mentions: The levels of nTregs in peripheral blood of MS patients and HC, are shown in Figure 1. nTreg levels were lower in acute-phase patients with no treatment (AP-noRx) or under methylprednisolone (AP-MP), compared to all other patient groups and HC. nTregs were restored to normal levels in acute-phase patients under interferon β treatment (AP-IFN), whereas patients in remission under no treatment (Rem-noRx) showed intermediate levels between HC and AP-noRx. Patients in remission under natalizumab (Rem-NATA) had nTreg levels comparable to those of AP-noRx and AP-MP patients (Figure 1D).


Regulatory Cell Populations in Relapsing-Remitting Multiple Sclerosis (RRMS) Patients: Effect of Disease Activity and Treatment Regimens
Flow cytometric analysis to determine CD4+CD25+Foxp3+ RC levels in human peripheral blood. A representative analysis is shown for one healthy control (A1–C1) and one relapsing remitting MS (RRMS) patient (A2–C2). The white blood cells (WBC) were gated on lymphocytes, based on forward and side light scatter (A1,A2) and analyzed for CD4 and CD25 expression (B1,B2); The double positive cells were analyzed further for Foxp3 expression (C1,C2); 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). AP-noRx (n = 13), patients in the acute phase of the disease without treatment; AP-MP (n = 17), patients in the acute phase under treatment with methylprednisolone; AP-IFN (n = 12), patients in the acute phase under treatment with interferon β; Rem-noRx (n = 15), patients in remission receiving no treatment; Rem-NATA (n = 26), patients in remission under treatment with natalizumab. * p = 0.04, ** p = 0.01, *** p = 0.002.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-17-01398-f001: Flow cytometric analysis to determine CD4+CD25+Foxp3+ RC levels in human peripheral blood. A representative analysis is shown for one healthy control (A1–C1) and one relapsing remitting MS (RRMS) patient (A2–C2). The white blood cells (WBC) were gated on lymphocytes, based on forward and side light scatter (A1,A2) and analyzed for CD4 and CD25 expression (B1,B2); The double positive cells were analyzed further for Foxp3 expression (C1,C2); 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). AP-noRx (n = 13), patients in the acute phase of the disease without treatment; AP-MP (n = 17), patients in the acute phase under treatment with methylprednisolone; AP-IFN (n = 12), patients in the acute phase under treatment with interferon β; Rem-noRx (n = 15), patients in remission receiving no treatment; Rem-NATA (n = 26), patients in remission under treatment with natalizumab. * p = 0.04, ** p = 0.01, *** p = 0.002.
Mentions: The levels of nTregs in peripheral blood of MS patients and HC, are shown in Figure 1. nTreg levels were lower in acute-phase patients with no treatment (AP-noRx) or under methylprednisolone (AP-MP), compared to all other patient groups and HC. nTregs were restored to normal levels in acute-phase patients under interferon β treatment (AP-IFN), whereas patients in remission under no treatment (Rem-noRx) showed intermediate levels between HC and AP-noRx. Patients in remission under natalizumab (Rem-NATA) had nTreg levels comparable to those of AP-noRx and AP-MP patients (Figure 1D).

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.


Related in: MedlinePlus