Adjuvant selection regulates gut migration and phenotypic diversity of antigen-specific CD4+ T cells following parenteral immunization.

Infectious diarrheal diseases are the second leading cause of death in children under 5 years, making vaccines against these diseases a high priority. It is known that certain vaccine adjuvants, chiefly bacterial ADP-ribosylating enterotoxins, can induce mucosal antibodies when delivered parenterally. Based on this, we reasoned vaccine-specific mucosal cellular immunity could be induced via parenteral immunization with these adjuvants. Here, we show that, in contrast to the Toll-like receptor-9 agonist CpG, intradermal immunization with non-toxic double-mutant heat-labile toxin (dmLT) from enterotoxigenic Escherichia coli drove endogenous, antigen-specific CD4+ T cells to expand and upregulate the gut-homing integrin α4ß7. This was followed by T-cell migration into gut-draining lymph nodes and both small and large intestines. We also found that dmLT produces a balanced T-helper 1 and 17 (Th1 and Th17) response, whereas T cells from CpG immunized mice were predominantly Th1. Immunization with dmLT preferentially engaged CD103+ dendritic cells (DCs) compared with CpG, and mice deficient in CD103+ DCs were unable to fully license antigen-specific T-cell migration to the intestinal mucosae following parenteral immunization. This work has the potential to redirect the design of existing and next generation vaccines to elicit pathogen-specific immunity in the intestinal tract with non-mucosal immunization.

Relationship between arterial thrombosis and neutralization of a polyethylene ionomer.

The influence of three levels of sodium neutralization of an ethylene/methacrylic acid copolymer on in vivo blood compatibility was studied in a canine arterial model. Effects due to sample fabrication methods (thermal pressing versus solution casting) were also monitored. Sodium content, sodium release, hydrogen dissociation, and localization of anionic groups were noted. Polymer surface energy, surface morphology, water uptake, and thermal properties were characterized. Material characterization and in vivo implantation disclose the following: 1) Thermal pressing generated oxidation degradation products that decreased in vivo blood compatibility. Solution-cast samples adhered and activated fewer blood elements. 2) Platelets and leukocytes were sensitive to differences in shear rate in the carotid and femoral arteries, with the femoral site tending toward higher shear, more platelet deposition and fewer leukocytes. 3) The surface properties of the polyethylene control, 0% Na, and 50% Na samples tended to be similar. These properties were different from the 100% Na sample. The 100% Na ionomer was more hydrophilic, had a higher polar component for its surface energy, and was unique in exhibiting discrete ionic clusters 1-10 microns in diameter on its surface. 4) These differences were manifested in vivo by platelet activation and thrombus development on the polyethylene, 0% Na, and 50% Na implants, while the 100% Na implant surfaces were predominantly covered by singly adherent, unactivated platelets. 5) It is proposed that the improvement in biocompatibility for the 100% Na ionomer is due to the cluster development in the neutralized methacrylic component and that either directly, or through appropriate protein adsorption and/or conformational adjustment to the cluster regions, platelets are not activated and do not initiate the coagulation mechanism.

Vascular graft sealants.

Although the use of vascular graft sealants is primarily intended to decrease the porosity of grafts upon implantation, sealants also may serve to passivate polymeric surfaces or act as a temporary scaffold for cell attachment and subsequent graft healing. The safety and efficacy of vascular grafts sealed with albumin, collagen, and gelatin have been established through many preclinical and clinical studies. Such products have been used in clinical practice since the mid-1980s. Any differences in the long-term human healing response to these sealants and the clinical relevance of such differences can be assessed only as the time of implantation increases and the data are evaluated.

Tissue culture of fetal rat islets: corticosterone promotes D cell maintenance and function.

Fetal pancreatic islets were cultured using a recently described technique (1). After 1 day in culture, half of the plates were continued in control medium and half were grown in identical medium supplemented with corticosterone (0.1 microgram/ml). Media were renewed daily, and culture was continued for a total of 8 days. Insulin, glucagon, and somatostatin contents in the media were determined daily. These hormones were also estimated in the tissue at the time of plating, and after 1 and 8 days in culture. Islets were fixed on day 8, and the cells containing each of these hormones were identified by immunocytochemical staining. Corticosterone supplementation of the medium resulted in a 3-fold increase in the somatostatin concentration of the medium. The insulin and glucagon contents of the supplemented medium were slightly reduced. On day 8, there was no difference in the insulin content of the cultured tissue regardless of medium. The glucagon and somatostatin contents of the tissue grown in the steroid-supplemented medium were greater (1.8- and 3.1-fold, respectively) than those of the tissue grown in control medium. D cells were rare in the islets grown in control medium volume density, 0.4%, but were more numerous in the islets maintained in supplemented medium (2.2%). Islets grown in corticosterone-supplemented medium lacked an insulin secretory response to 22 mM glucose. These findings indicate that the volume densities of the cells within the islets can be altered during an 8-day period in culture, suggesting that nutritional and other requirements of the individual subpopulations of islet cells may be different. In addition, corticosterone may prevent the maturation of the secretory responsiveness of cultured B cells to glucose.