Cytokine balance and lipid antigen presentation in the NOD mouse pancreas during development of insulitis.

The role of cytokine balance and lipid antigen presentation in the development of diabetes was studied using immunohistochemistry of cytokines in the pancreas of non-obese diabetic mice (NOD) and BALB/c mice at various ages. In both the NOD and BALB/c mice, interleukin 10 (IL-10) was expressed in the islets. IL-10 was also present in the epithelial cells of the exocrine tissue in both strains. In the NOD mice, IL-10 disappeared from both the islets and the exocrine tissue at 16 weeks of age. At this age, IL-10 was still present in the islets and exocrine tissue of the BALB/c pancreata. IL-10 was not present in the pancreata of diabetic NOD mice. IL-6 first appeared in the pancreas at 10 weeks of age and disappeared at the age of 16 weeks in both NOD and BALB/c mice. It was present in the endothelial cells. Neither the pancreata of normal BALB/c mice nor NOD mice at 2-16 weeks of age contained tumor necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma), IL-4, or IL-12. At 8 weeks of age, a few IL-2+ cells were found in the pancreas of one of three NOD mice. CD1d was already present in both strains at 2 weeks of age but disappeared from the NOD mice at 16 weeks of age. CD1d localized to walls of tubular structures probably representing collecting tubules. These results suggest that in the NOD mice the disappearance of the T(H0), T(H1), and T(H2) responses inhibiting IL-10 from the islets at the age of 16 weeks may trigger the final stage of the immune response leading to overt diabetes. The simultaneous disappearance of CD1d suggests that activation of immune responses against lipid antigens does not play a role in this stage of the disease.

The role of lipid antigen presentation, cytokine balance, and major histocompatibility complex in a novel murine model of adoptive transfer of insulitis.

After adoptive transfer of insulitis from nonobese diabetic (NOD) mice, leukocytes accumulate in the pancreas of SCID/SCID and NOD/SCID mice. These cells express classical antigen-presenting molecules and costimulators of T-cell activation and adhesion molecules involved in homing. The aim of the present study was to study the expression of cytokines involved in regulation of the T(H1)/T(H2) balance by these cells, the role of lipid antigen presentation in the local immune system activation in the pancreas during onset of insulitis, and the role of major histocompatibility complex in this process. Splenocytes from NOD and BALB/c mice were injected intraperitoneally to SCID/SCID and NOD/SCID mice. Sections from the pancreata of these injected mice were stained for cytokines (tumor necrosis factor alpha [TNF-alpha], interferon gamma [IFN-gamma], CD1d, interleukin 2 [IL-2], IL-4, IL-6, IL-10, and IL-12). Some SCID/SCID and NOD/SCID mice injected with NOD splenocytes developed a severe disease. IL-10 was expressed in almost all the animals: in exocrine pancreas, large groups of infiltrating lymphocytes, endothelia of blood vessels, pancreatic islets, and interstitial tissue. CD1d was found in most of the mice: in the endothelia of collecting ducts and blood vessels of the pancreas, lymphocytic infiltrates, interstitial tissue, septae, islets, and a pancreatic lymph node. TNF-alpha was expressed notably more often in the pancreata of NOD/SCID than SCID/SCID mice. It was found between pancreatic lobules, in the epithelia of collecting ducts, endothelia of blood vessels, islets, capillaries, infiltrates, and septae. IL-6 was expressed more in the SCID/SCID than in the NOD/SCID mice. It was seen in infiltrates, walls of blood vessels, around islets, and in connective tissue. IFN-gamma was found only in the pancreata of SCID/SCID and NOD/SCID mice injected with NOD splenocytes. The expression of IL-2 and IL-12 was very scarce. IL-4 was not expressed at all. The present study clearly shows that antigen presentation has a role in the development of autoimmune diseases after adoptive transfer of splenocytes from diseased mice to intact ones and that IL-10 may have a central role in the control of the disease process.

Gait Speed and Handgrip Strength as Predictors of Incident Disability in Mexican Older Adults.

Physical performance tests are associated with different adverse outcomes in older people. The objective of this study was to test the association between handgrip strength and gait speed with incident disability in community-dwelling, well-functioning, Mexican older adults (age ≥70 years). Incident disability was defined as the onset of any difficulty in basic or instrumental activities of daily living. Of a total of 133 participants, 52.6% (n=70) experienced incident disability during one year of follow-up. Significant associations of handgrip strength (odds ratio [OR] 0.96, 95% confidence interval [95%CI] 0.93-0.99) and gait speed (OR 0.27, 95%CI 0.07-0.99) with incident disability were reported. The inclusion of covariates in the models reduced the statistical significance of the associations without substantially modifying the magnitude of them. Handgrip strength and gait speed are independently associated with incident disability in Mexican older adults.

Testicular-associated immune deviation: flushing of the testicular lymph sinusoids induces immunosuppression and inhibits formation of EAE in SJL mice.

Injection of antigen into the testis has been previously proved to induce systemic tolerance in rats. Testicular-associated immune deviation (TAID) has thus far been induced and studied only in the rat and the present study was planned to study if TAID could be induced in mice as well. In addition, it was studied if TAID is organ-specific. Mouse spinal cord homogenate (MSCH), as well as phosphate-buffered saline (PBS), was injected into the testes of SJL and BALB/c male mice before the induction of experimental allergic encephalomyelitis into the animals. The control animals received MSCH intramuscularly into the hamstring muscles. The animals were followed and graded daily for symptoms attending the next 30 days. In the SJL strain, mice treated with an intratesticular (i.t.) MSCH injection prior to the induction of experimental autoimmune encephalomyelitis (EAE) had the shortest duration of symptoms and the longest time to the onset of the first symptoms. In addition, the mice injected i.t. with PBS had as mild symptoms as those injected with MSCH. There was a statistically significant difference, however, between the groups injected either with MSCH or PBS intratesticularly. In general, mice treated with an intramuscular injection of MSCH got sick first, and had the most severe symptoms for the longest duration of time. In the case of the BALB/c mice, there were no statistical differences between the groups investigated. It is concluded that TAID is a testis- and strain-specific phenomenon in the mouse, and not specific to the rat. In addition, i.t. injection of PBS is just as effective in creating tolerance against EAE as i.t. injection of MSCH in the SJL mice.

Expression of cyclooxygenase-2 in intestinal goblet cells of pre-diabetic NOD mice.

Cyclooxygenase, the rate-limiting enzyme in prostaglandin synthesis, is expressed in constitutive (COX-1) and inducible (COX-2) isoforms. The COX-2 has been proposed to be involved in development of autoimmune type 1 diabetes (T1D). We examined COX-2 expression in the gut-associated lymphoid tissue (GALT), and found COX-2 was strongly expressed in goblet cells of non-obese diabetic (NOD) mice at the apical villi at the age of 2.5 weeks, clearly before the onset of insulitis, while the expression in the control BALB/c mice was weak or absent at all ages (P < 0.001). Lipopolysaccharide (LPS) given intraperitoneally slightly increased COX-2 expression in the goblet cells and epithelium of both NOD and BALB/c mice. High-resolution confocal microscopy showed that the surroundings of the goblet cells contained no COX-2, implying that the enzyme is synthesized by the goblet cells. The COX-2 is secreted from goblet cells into the intestinal lumen along with mucins. The COX-2 concentration in the goblet cell of BALB/c and especially of NOD mice was markedly higher than that in the intraepithelial lymphocytes or lamina propria macrophages. High mucin COX-2 from goblet cells may increase luminal prostaglandin synthesis, alter epithelial permeability, modulate intestinal immune responses and modify functional properties of the lymphocytes in the GALT, which all may be important for the initiation of the autoimmune phenomenon in the NOD mice.