Dopaminergic Receptors on CD4+ T Naive and Memory Lymphocytes Correlate with Motor Impairment in Patients with Parkinson's Disease

Natasa Kustrimovic, Emanuela Rasini, Massimiliano Legnaro, Raffaella Bombelli, Iva Aleksic, Fabio Blandini, Cristoforo Comi, Marco Mauri, Brigida Minafra, Giulio Riboldazzi, Vanesa Sanchez-Guajardo, Franca Marino, Marco Cosentino, Natasa Kustrimovic, Emanuela Rasini, Massimiliano Legnaro, Raffaella Bombelli, Iva Aleksic, Fabio Blandini, Cristoforo Comi, Marco Mauri, Brigida Minafra, Giulio Riboldazzi, Vanesa Sanchez-Guajardo, Franca Marino, Marco Cosentino

Abstract

Parkinson's disease (PD) is characterized by loss of dopaminergic neurons in substantia nigra pars compacta, α-synuclein (α-syn)-rich intraneuronal inclusions (Lewy bodies), and microglial activation. Emerging evidence suggests that CD4+ T lymphocytes contribute to neuroinflammation in PD. Since the mainstay of PD treatment is dopaminergic substitution therapy and dopamine is an established transmitter connecting nervous and immune systems, we examined CD4+ T naive and memory lymphocytes in PD patients and in healthy subjects (HS), with specific regard to dopaminergic receptor (DR) expression. In addition, the in vitro effects of α-syn were assessed on CD4+ T naive and memory cells. Results showed extensive association between DR expression in T lymphocytes and motor dysfunction, as assessed by UPDRS Part III score. In total and CD4+ T naive cells expression of D1-like DR decrease, while in T memory cells D2-like DR increase with increasing score. In vitro, α-syn increased CD4+ T memory cells, possibly to a different extent in PD patients and in HS, and affected DR expression with cell subset-specific patterns. The present results support the involvement of peripheral adaptive immunity in PD, and may contribute to develop novel immunotherapies for PD, as well as to better use of current dopaminergic antiparkinson drugs.

Figures

Figure 1. CD4+ T naive and memory…
Figure 1. CD4+ T naive and memory cells in HS and PD patients.
Cells are shown as absolute numbers (panels a,b) and as percentage of total CD4+ T cells (c,d). Data are shown as medians with 25°–75° percentiles (boxes) and min-max values (whiskers). Comparisons are shown between HS and PD patients as a whole (a,c) and between drug naive (PD-dn) and drug treated (PD-dt) patients (b,d). Differences were analyzed by means of two-tailed Student’s t test or by Mann-Whitney test, as appropriate. P values less than 0.05 are indicated in the graphs.
Figure 2. DR expression on CD4+ T…
Figure 2. DR expression on CD4+ T cells from HS and from PD patients.
DR expression is shown as mRNA levels (panels a,b) and as protein expression on the membranes of CD4+ T cells, expressed as absolute numbers of DR+ cells (c,d) and as percentage of total CD4+ T cells (e,f). Comparisons are shown between HS and PD patients as a whole (a,c,e) and between drug naive (PD-dn) and drug treated (PD-dt) patients (b,d,f). Data are shown as medians with 25°–75° percentiles (boxes) and min-max values (whiskers). Differences were analyzed by means of two-tailed Student’s t test or by Mann-Whitney test, as appropriate. P values less than 0.05 are indicated in the graphs.
Figure 3. DR expression on CD4+ T…
Figure 3. DR expression on CD4+ T naive cells from HS and from PD patients.
DR+ cells are shown as absolute numbers (panels a,b) and as percentage of total CD4+ cells (c,d). Data are shown as medians with 25°–75° percentiles (boxes) and min-max values (whiskers). Comparisons are shown between HS and PD patients as a whole (a,c) and between drug naive (PD-dn) and drug treated (PD-dt) patients (b,d). Differences were analyzed by means of two-tailed Student’s t test or by Mann-Whitney test, as appropriate. P values less than 0.05 are indicated in the graphs.
Figure 4. DR expression on CD4+ T…
Figure 4. DR expression on CD4+ T cells and UPDRS-III score.
DR expression is shown as mRNA levels (panel a) and as protein expression on the membranes of CD4+ T cells, expressed as absolute numbers of DR+ cells (b) and as percentage of total CD4+ T cells (c) Data are medians with 25°–75° percentiles (boxes) and min-max values (whiskers). Differences in DR expression between HS and PD patients were analyzed by parametric ANOVA or Kruskal-Wallis nonparametric ANOVA, as appropriate, with either Holm-Sidak’s or Dunn’s adjustments for multiple comparisons, where * = P < 0.05 and ** = P < 0.01. Trend analysis in PD patients was performed by ANOVA post test for linear trend.
Figure 5. DR expression in CD4+ T…
Figure 5. DR expression in CD4+ T naïve, TCM and TEM and UPDRS-III score.
DR expression is shown as protein expression on the membranes of CD4+ T naïve (left), TCM (center) and TEM (right) cells, expressed as absolute numbers of DR+ cells. Data are medians with 25°–75° percentiles (boxes) and min-max values (whiskers). Differences between DR levels in HS and in PD patients were analyzed by parametric ANOVA or Kruskal-Wallis nonparametric ANOVA, with either Holm-Sidak’s or Dunn’s adjustments for multiple comparisons, where * = P < 0.05 and ** = P < 0.01. Trend analysis in PD patients was performed by ANOVA post test for linear trend.
Figure 6. Effect of TTd, monomeric, and…
Figure 6. Effect of TTd, monomeric, and fibrillar α-syn on the frequency of CD4+ T naive and memory subsets.
Panel (a) Effects of TTd (left), monomeric (middle), and fibrillar α-syn (right) in cells from HS (open columns) and PD patients (hatched columns). Data are expressed as percentage variation with respect to control conditions (without TTd or α-syn), and are means ± SEM of n = 6–8 separate experiments each performed in duplicate. * = P < 0.05 and ** = P < 0.01 vs. control conditions, and # = P < 0.01 vs. HS. Panel (b) monomeric (hatched columns), and fibrillar α-syn (shaded columns) on DR expression in T naive (left), TCM (center) and TEM cells (right) from 5 HS. Data are means ± SEM. * = P < 0.05 and ** = P < 0.01 vs control (open columns); # = P < 0.01 vs monomeric α-syn.

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