Transfusion of Nonobese Diabetic Mice with Allogeneic Newborn Blood Ameliorates Autoimmune Diabetes and Modifies the Expression of Selected Immune Response Genes

Abstract

Although allogeneic bone marrow transplantation has been shown to prevent autoimmune diabetes in heavily irradiated NOD mice, similar procedure is not suitable for the treatment of patients with type 1 diabetes because of associated severe side effects. Therefore, we evaluated whether mouse newborn blood, equivalent to human umbilical cord blood, could be used for diabetes prevention without recipient preconditioning. To test this hypothesis, unconditioned, pre-diabetic female NOD mice were given a single injection of whole newborn blood derived from the allogeneic, diabetes-resistant mouse strain, C57BL/6. Transfusion of allogeneic newborn but not adult blood prevented diabetes incidence in a majority of treated mice for a prolonged period of time. This was accompanied by the release of insulin in response to a challenge with glucose. Invasive cellular infiltration of islets was also substantially reduced in these mice. Although newborn blood transfusion induced low level of hematopoietic microchimerism, it did not strictly correlate with amelioration of diabetes. Induction of genes implicated in diabetes such as, Il18, Tnfa, and Inos but not Il4, Il17 or Ifng was repressed in splenocytes derived from protected mice. Notably, expression of the transcription factor Tbet/Tbx21 but not Gata3 or Rorgt was upregulated in protected mice. These data indicate that allogeneic newborn blood transfusion can prevent diabetes in NOD mice associated with modulation of selected cytokine genes implicated in diabetes manifestation. The data presented herein provide the proof of principle for the utility of allogeneic umbilical cord blood transfusion to treat patients with autoimmune diabetes.

Transcriptome Analysis of Epigenetically Modulated Genome Indicates Signature Genes in Manifestation of Type 1 Diabetes and Its Prevention in NOD Mice

“…Non-fasting blood glucose levels were monitored weekly and >250 mg/dL for two consecutive weeks were considered diabetic [8], [22]. Whereas overtly diabetic mice were killed between 24 and 28 wk of age, TSA-treated and cured mice were killed between 28 and 34 wk of age and spleens and pancreata harvested.…”

“…Whereas overtly diabetic mice were killed between 24 and 28 wk of age, TSA-treated and cured mice were killed between 28 and 34 wk of age and spleens and pancreata harvested. Histological analysis of paraffin embedded pancreata was performed as described earlier [22]. Each NOD.…”

“…TSA-treated mice that remained diabetic were not included in the analysis. Total RNA was extracted from individual spleens after lysing in TRIzol, as described [8], [22]. RNA from 4 to 6 mice per group was pooled to minimize the expression bias.…”

“…Total RNA was extracted from un-induced splenocytes and those stimulated with immobilized anti-CD3 antibody for 18 h as described earlier [8], [22]. Pancreata were stored in RNALater (Life Technologies, Grand Island, NY) at −80°C and RNA was extracted from pancreata dissociated in TRIzol.…”

Y-chromosome DNA Is Present in the Blood of Female Dogs Suggesting the Presence of Fetal Microchimerism

“…Unraveling the function of these microchimeric cells is imperative to provide insight into their role in health and disease. Currently, mouse models have identified benefits of induced FMC in chronic disease risk [11]. A naturally occurring, large animal model with risk for spontaneous diseases that frequently affect humans and which have a high frequency of FMC will allow hypothesis testing in a more realistic setting.…”