Adoptive Transfer of Diabetes in BB Rats Induced by CD4 T Lymphocytes

Abstract

Unseparated splenocytes (SPCs) or purified SPC subsets from diabetes-prone BB (BBdp) or diabetic BB (BBd) rats were activated in vitro with either phorbol myristate acetate (PMA) and ionomycin (I) or concanavalin A (ConA). Such activated SPCs were then injected intravenously into 30-day-old BBdp rats, and their capacity to induce adoptive transfer (AT) of diabetes was studied. The proliferative response in vitro of BBd unseparated SPCs or purified W3/13+ SPCs (i.e., T lymphocytes + large granular lymphocytes) to PMA + I far exceeded that of ConA, resulting in mean stimulation indices of 68 and 112 (PMA + I) and 1.9 and 30 (ConA). The incidence of AT was similar when equal numbers of unseparated SPCs from the same BBd donor were injected after activation by either PMA + I + interleukin 2 (PII) or ConA (57 vs. 50%, respectively); however, injection of PII-activated and macrophage-depleted W3/13+ SPCs from BBd animals resulted in a significantly higher incidence of AT (90%, P less than 0.05). As few as 0.5 x 10(6) PII-activated W3/13+ SPCs were sufficient to induce AT. Sixteen percent of recipients developed diabetes after injection of activated W3/13+ cells from 40-day-old BBdp donors. To determine which W3/13+ cells might mediate such transfer, purified and PII-preactivated CD4 T lymphocytes from BBd rats were injected, and they succeeded in AT in 44% of the recipients. Preactivated BBd B lymphocytes were unable to induce AT. Although a possible role for large granular lymphocytes cannot be excluded, the results demonstrate that in the BB rat, the beta-cell destruction can be induced by CD4 T lymphocytes.

Prevention of spontaneous immune-mediated diabetes development in the LEW.1AR1-iddm rat by selective CD8+ T cell transfer is associated with a cytokine shift in the pancreas-draining lymph nodes

“…Thus, a clear autoaggressive potential of CD4 + T cells was seen, in line with several studies on BB-DP rats and NOD mice [7][8][9][10][11][12]. The predominant role of CD4 + T cells has been previously demonstrated by selective transfer of CD4 + or CD8 + T cells in immunodeficient NOD strains, whereas CD8 + T cell subsets were not diabetogenic [7,16].…”

“…The only difference between the LEW.1AR1-iddm rat and the background strain is a mutation on chromosome 1 within Iddm8 (RNO1q43-51) [4,5]. Adoptive transfer of T cells from diabetic LEW.1AR1-iddm rats to prediabetic LEW.1AR1-iddm rats significantly increased the incidence of diabetes [6] in agreement with studies on NOD mice and BB-DP rats [7][8][9][10][11][12]. The autoreactive potential can be transferred through immune cells from diabetic LEW.1AR1-iddm rats into athymic nude LEW.1AR1-Whn rnu animals, but not to the diabetes-resistant LEW.1AR1 background strain [6].…”

Low Dose Poly I:C Prevents Diabetes in the Diabetes Prone BB Rat

“…Effector cells such as T cells [16] and NK cells [17] are reported to play an important role in the development of diabetes in the BB rat. Several strategies have been attempted to suppress or eliminate effector cells, and in turn prevent diabetes.…”

“…Three micrograms of splenic total RNAs were converted to cDNAs using Superscript II (Life Technologies, Gaithersburg, MD, USA) and oligo (dT) [12][13][14][15][16][17][18] (Life Technologies) in 20 l of the reaction mixture at 42°C for 1 h. After the synthesis of cDNA, PCR was run using specific primers for IFN-, IL-2, IL-4, IL-10 and TNF as previously described [15]. Hypoxanthine phosphoribosyltransferase (HPRT) was used as an internal standard.…”

Impaired Post-Thymic Development of Regulatory CD4+25+ T Cells Contributes to Diabetes Pathogenesis in BB Rats

“…The disease is polygenic, T cell-mediated, and affects most males and females around puberty (1)(2)(3)(4)(5)(6)(7). Two groups have recently reported the positional cloning of the BB rat diabetes susceptibility gene Iddm2/Lyp located on chromosome 4 (8,9).…”