Cord Blood Cytokine Profile Detection in Neonates with T1D Parents – Monitoring of Cellular Auto‐reactivity Using Protein Microarray

Abstract

Type 1 diabetes (T1D) is a great medical challenge and its incidence rises rapidly. T lymphocytes and their cytokine production are supposed to play a major role in T1D development. So far, there is no potent tool to recognize the early signs of cellular auto‐reactivity which leads to β‐cell damage. The naïve immune system of the newborn (not yet influenced by external factors) can be used as an important model for T1D pathogenesis studies. Cord blood samples of 22 healthy neonates born at term to a diabetic parent (T1DR) and 15 newborns with no family history of any autoimmune disease (controls) were collected. Determination of 23 cytokines was performed before and after the stimulation with diabetogenic autoantigens using protein microarray. We observed lower basal production of all detected cytokines in the T1DR group – granulocyte/macrophage colony‐stimulating factor (GM‐CSF) (P = 0.025), growth regulated protein (GRO) (P = 0.002), GRO‐α (P = 0.027), interleukin (IL)‐1‐α (P = 0.051), IL‐3 (P = 0.008), IL‐7 (P = 0.027), IL‐8 (P = 0.042), monocyte chemoattractant proteins (MCP)‐3 (P = 0.022), monokine‐induced by IFN‐γ (MIG) (P = 0.034) and regulated upon activation normal T‐cell express sequence (RANTES) (P = 0.004). Exclusively lower post‐stimulative levels of G‐CSF (P = 0.030) and GRO‐α (P = 0.04) were observed in controls in comparison with the basal levels. A significant post‐stimulative decrease in G‐CSF (P = 0.030) and MCP‐2 (P = 0.009) levels was observed in controls in comparison with T1DR neonates. We also observed the interesting impact of the risky genotype on the protein microarray results. Protein microarray seems to be a useful tool to characterize a risk pattern of the immune response for T1D also in newborns.

Differential gene expression pattern of diabetic rat retinas after intravitreal injection of erythropoietin

“…After EPO treatment, the expression of CD80 and CD86 was upregulated (data not shown). Lower basal production of IL-7 was detected in DR group by the protein microarray, 30 and in our study, the mRNA expression of IL-7 was downregulated, especially in the mid-late stage, while it was upregulated after intravitreal EPO treatment. Col1a2 (procollagen, type I, alpha 2) was significantly downregulated in the insulin-deficient, hyperglycaemic diabetic animals, and in our experiment, the expression of Col1a2 was downregulated at intermediate and late stages of diabetic rats, and intravitreal EPO can reversed the result (data not shown).…”

Influence of Maternal Hyperglycaemia on Cord Blood Mononuclear Cells in Response to Diabetes‐associated Autoantigens

“…The first one is the influence of maternal 'autoimmune'environment and the second one is the influence of maternal diabetes in itself (mainly maternal hyperglycaemia) on the foetal immune reactivity against autoantigens. In the present as well as in our previous study [9], we observed that, under certain conditions, cord blood mononuclear cells are able to react by Th1 predominance to a mixture of diabetogenic peptides. Our previous study focused on Th1 predominance after specific stimulation observed in children carrying alleles for HLA class II molecules with considerable risk for development of T1D [9].…”

“…In the present as well as in our previous study [9], we observed that, under certain conditions, cord blood mononuclear cells are able to react by Th1 predominance to a mixture of diabetogenic peptides. Our previous study focused on Th1 predominance after specific stimulation observed in children carrying alleles for HLA class II molecules with considerable risk for development of T1D [9]. In the present study, children with T1D father or T1D mother with perfect diabetes compensation during the 3rd trimester of pregnancy were studied.…”

“…We have previously shown that the cytokine‐ and chemokine‐spectrum differs between children born to T1D mothers and mothers without T1D [9]. In the present study, we expand our cohort to include also children of T1D father as well as of children born to mothers with other forms of diabetes; gestational and type 2 diabetes (T2D).…”

Autoimmunity: limited progress for the patient, despite decades of research

“…For example, a similar search in MedLine indicated that 40 papers on autoimmunity were published over the past two and one‐half years [11–50]. While 19 of these deal primarily with general immunology [11–26] genetics [27] and technologies [28, 29], the remaining 21 papers [30–50] focus on specific autoimmune diseases. Not surprising, the more popular topics have been multiple sclerosis/experimental autoimmune encephalomyelitis (six manuscripts) [30–35] diabetes/Addison’s disease (four manuscripts) [36–39] and rheumatic diseases/collagen‐induced arthritis (four manuscripts) [40–43].…”