Małgorzata Bilińska

Association of the HLA‐G gene polymorphism with multiple sclerosis in a Polish population

In this study, three polymorphic sites in the HLA‐G gene: −725C>G>T, −716T>G and 14bpindel were genotyped. Significant differences were found between patients and controls in the alleles and genotypes for −725C>G>T and in three‐point haplotypes. We observed also a significant difference in the age of disease onset between patients positive and negative for 14bpins. The results suggest that single nucleotide polymorphisms in the promoter of the HLA‐G gene (mainly −725C>G>T), and 14bpindel, or some genetic marker in tight linkage disequilibrium with them are associated with multiple sclerosis.

6.7-kbp deletion in LILRA3 (ILT6) gene is associated with later onset of the multiple sclerosis in a Polish population

Recently published studies have implicated the deletion polymorphism in LILRA3 gene, as being associated with multiple sclerosis (MS). A total of 309 patients diagnosed with MS and 379 unrelated healthy volunteers were typed for 6.7-kbp deletion in LILRA3 gene. Simultaneously, presence or absence of HLA-DRB1(∗)1501 allele was established to assess the possibility of interaction between LILRA3 deletion and HLA-DRB1(∗)1501 status. In contrast to previous reports, we did not find any association of LILRA3 deletion with MS susceptibility. Also, the HLA-DRB1(∗)1501 stratification analysis showed no LILRA3 association with the disease. However, we observed that patients negative for the deletion may begin to suffer from MS significantly earlier than patients who are positive (p = 0.014). Similarly to the most European populations we found significantly higher frequency of HLA-DRB1(∗)1501 allele in cases than we found in controls (27.0% vs. 12.5%; p < 0.0001, OR = 2.6, 95%CI = 1.96-3.42).

Different patterns of activation markers expression and CD4+ T-cell responses to ex vivo stimulation in patients with clinically quiescent multiple sclerosis (MS)

Patients with relapsing-remitting (RR) and secondary progressive (SP) forms of multiple sclerosis (MS), although in long-term clinical remission, showed different patterns of increased expressions of the activation markers: CD69, CD40L, and both membrane/surface and cytoplasmic CTLA-4 (mCTLA-4 and cCTLA-4, respectively) in freshly isolated peripheral blood (PB) CD4+ T cells compared with controls. Also observed were dysregulated responses to ex vivo stimulation in both groups of MS patients accompanied by increased IFN-gamma synthesis. Our findings may suggest that the mechanisms leading to each clinical form of the disease may be heterogeneous.

ALCAM and CD6 — multiple sclerosis risk factors

ALCAM and CD6 may play an important role in the pathogenesis of multiple sclerosis (MS), since they are involved in the transmigration of leukocytes across the blood-brain barrier. In this study, we confirmed our previous findings about the association of the ALCAM gene with risk, development and progression of MS. Additionally, we showed that in the case of the CD6 gene (encoding receptor of ALCAM) not only polymorphisms but also mRNA expression level are associated with MS. Our analysis revealed that the risk of the disease for AA individuals in rs12360861 was almost 3.0-fold lower in comparison to GG individuals (OR=0.34; CI95%=0.12; 0.81). Moreover, we observed lower expression of CD6 mRNA in patients than in healthy individuals (T(2)2,74=6.678; p=0.002).

ALCAM — Novel multiple sclerosis locus interfering with HLA-DRB1*1501

Activated leukocyte cell adhesion molecule (ALCAM) is a molecule involved in leukocyte migration across the blood-brain barrier which is a key stage in multiple sclerosis (MS) pathogenesis. The present study is the first to report evidence of the association of rs6437585 ALCAM polymorphism with risk and progression of MS. Our investigation revealed that rs6437585CT individuals had higher risk of MS (OR=2.34; 95%CI=1.22-4.51; P=0.011) and over 2 years earlier age of onset (95%CI=0.16-4.41, P=0.036). Moreover, we demonstrated that two ALCAM polymorphisms, rs11559013 and rs34926152, although not associated with MS itself, modify HLA-DRB1*1501 effect. Results obtained from logistic regression analysis showed five-fold lower risk for MS for both rs11559013GA/HLA-DRB1*1501+ and rs34926152GT/HLA-DRB1*1501+ individuals. This observations may suggest protective role against MS for both rs11559013GA and rs34926152GT genotypes in HLA-DRB1*1501 positive individuals.

Polymorphisms in CD28, CTLA-4, CD80 and CD86 genes may influence the risk of multiple sclerosis and its age of onset

CD28/CTLA-4–CD80/CD86 molecules play an important role in the regulation of T cells activation. Defects in proteins involved in this pathway may lead to the development of autoimmune diseases in which T cells are involved. In this case–control study (336 multiple sclerosis (MS) patients and 322 controls) we investigated the possible association of eleven polymorphisms in CD28, CTLA-4, CD80 and CD86 genes with susceptibility to MS and/or its progression. We also took into account HLA-DRB1*15:01 status. Moreover, this study aimed to determine the possible gene-gene interactions between examined SNPs associated with the susceptibility to MS and its outcome. Our investigation revealed that in HLA-DRB1*15:01 negative individuals, G allele in rs231775A NGof CTLA-4 gene was associatedwith higher risk ofmultiple sclerosis. Additionally, the association of rs2715267T NGof CD86 gene withMS susceptibilitywas detected. In details, carriers of G allele at this polymorphic site possessed higher risk of MS in comparison to TT homozygotes. On the other hand, the lower risk of MS was observed in individuals carrying A allele at the rs1599795T N A polymorphic site of CD80. Furthermore, the analysis revealed an interaction between three polymorphisms: rs3116496T N C (CD28), rs6641T N G (CD80) and rs17281995G N C (CD86), associated with the age of MS onset.

Investigation of gene–gene interactions between CD40 and CD40L in Polish multiple sclerosis patients

CD40-CD40L interaction is necessary for the activation of both humoral and cellular immune response and has been suggested to play a role in the pathogenesis of multiple sclerosis (MS). Therefore, we analyzed the combined influence of the CD40 and CD40L variants on MS susceptibility and progression on well-defined Polish population. Our investigation revealed that CT individuals in rs1883832 locus of CD40 possessed almost 1.5-fold higher risk for MS than CC individuals (OR = 1.44; 95%CI = 1.03-2.1; p = 0.032), while this risk for TT individuals was almost 2.5-fold higher (OR = 2.36; 95%CI = 1.19-4.78; p = 0.014). Moreover, for the first time, we observed the association of CD40 gene with MS development and progression. We observed that for the rs1883832CC individuals the age at diagnosis was on average 2 years lower than for the rs1883832CT and rs1883832TT individuals (CI95% = -3.69-(-0.29); p = 0.023). Additionally, we detected that individuals with TT and CT genotypes showed lower risk of developing secondary progressive course in comparison to those with CC genotype. For rs1883832TT individuals this risk was 4-fold lower (HR = 0.24; CI95% = 0.10-0.53; p = 0.00062). Despite the fact that CD40-CD40L pathway plays a key role in development of autoimmune diseases, we were not able to detect gene-gene interactions between CD40 and CD40L polymorphisms associated with multiple sclerosis.

MS risk allele rs1883832T is associated with decreased mRNA expression of CD40

CD40-CD40L interactions mediate T-dependent B cell response and efficient T cell priming. Therefore, genes encoding these molecules are attractive candidates for studies on autoimmune diseases, such as multiple sclerosis (MS), in which activated T and B cells are involved. Thus, we analyzed CD40 and CD40L mRNA expression in whole blood samples from MS patients and controls. Additionally, we examined the effect of three SNPs of CD40 (rs1883832C>T, rs11569343C>G, and rs752118C>T) and two SNPs of CD40L (rs3092923T>C and rs3092952A>G) on their mRNA expression. Our results showed that the rs1883832C>T SNP affects CD40 gene expression. Our analysis revealed that individuals possessing CT and TT genotypes (predisposing to MS) had decreased level of CD40 mRNA in comparison to those with CC. Moreover, we demonstrated the potential role of impaired CD40-CD40L interaction in developing of multiple sclerosis.

PD-1 gene polymorphic variation is linked with first symptom of disease and severity of relapsing-remitting form of MS

Multiple sclerosis (MS) is an inflammatory disease of the central nervous system (CNS), where inflammation, demyelination together with the axonopathy are the cardinal features on pathologic ground, with a combined genetic and environmental background. The associations of PD-1 single nucleotide polymorphisms (SNPs): PD-1.3 (in intron 4), PD-1.5 and PD-1.9 (both in exon 5) with clinical manifestation of MS in 479 south Polish individuals including 203 MS patients were analyzed. Presence of PD-1.5T allele was linked with the first manifestations of disease: diplopia and pyramidal signs - favored pyramidal signs but protected against of diplopia development. Farther, PD-1.3G/PD-1.5C/PD-1.9C haplotype significantly favored whereas GTC protected against diplopia. Besides, GTT haplotype strongly favored non-severe RRMS outcome and ATC haplotype was specific only for these MS patients. Our population-based case-control study, investigating selected three PD-1 SNPs: PD-1.3, PD-1.5 and PD-1.9, revealed that polymorphic variation may be rather disease-modifying than MS risk factor.

Profile of autonomic dysfunctions in patients with primary brain tumor and possible autoimmunity

OBJECTIVE Cerebral lesion due to different neurological conditions could be complicated by autonomic dysfunction, reported in the literature as a sympathetic hyperactivity. The mechanisms of dysautonomia still remains partial. The aim of the study was to assess the profile of autonomic dysfunction in patient with primary brain tumors, with attempt to estimate the additional factors in pathogenesis of dysautonomia. MATERIAL AND METHODS Neurological examinations, the Lows autonomic disorder questionnaire, electrophysiological autonomic tests (Heart Rate Variability test at rest and during deep breathing, spectral analysis of R-R intervals, sympathetic skin response test), studies of peripheral nerves, blood sampling collection for antibodies were done in 33 patients with recognized primary brain tumors. RESULTS The averaged Lows Questionnaire score in the patients group was significantly higher than in the controls, systolic blood pressure was increased, heart rate tended to be higher without significance, but heart rate variability was severe low, LF/HF ratio also tended to be higher in the patients group. In SSR test the amplitude of responses from hand and foot was significantly lower without changes in their latencies. We found changes in the electrophysiological tests of peripheral nerves, and positive anti-neural antibodies in 5 patients. CONCLUSIONS The results of the study indicated to the sympathetic nervous system hyperactivity in patients with primary brain tumors. Local brain lesion with high intracranial pressure, additional peripheral nerve damage probably in the course of autoimmunity, and direct influence of autoimmunity to the central part of autonomic nervous system are possible in the pathogenesis of dysautonomia.