X-ray crystal structure of HLA-DR4 (DRA*0101, DRB1*0401) complexed with a peptide from human collagen II.

Genetic predisposition to rheumatoid arthritis (RA) is linked to the MHC class II allele HLA-DR4. The charge of the amino acid at DRbeta71 in the peptide-binding site appears to be critical in discriminating DR molecules linked to increased disease susceptibility. We have determined the 2.5 A x-ray structure of the DR4 molecule with the strongest linkage to RA (DRB1*0401) complexed with a human collagen II peptide. Details of a predicted salt bridge between lysine DRbeta71 and aspartic acid at the P4 peptide position suggest how it may participate in both antigen binding and TCR activation. A model is proposed for the DR4 recognition of collagen II (261-273), an antigen immunodominant in human-transgenic mouse models of RA.

Do cytokines play a role in skeletal muscle ischemia and reperfusion?

Cytokines, interleukin-1 (IL-1) and tumor necrosis factor (TNF) are known to mediate host cell response to sepsis, trauma, and myocardial ischemia. We have previously found increased levels of IL-1 in the venous effluent during the reperfusion phase of skeletal muscle ischemia in a canine model. This study was done to evaluate whether TNF also played a role in skeletal muscle ischemia-reperfusion injury since IL-1 and TNF have inter-related functions. In twelve adult mongrel dogs (28-32 kg) one gracilis muscle was subjected to six hours of normothermic ischemia followed by normothermic reperfusion. The contralateral side served as a control and remained normally perfused throughout the experiment. Gracilis venous samples were collected at pre-ischemia and one hour of reperfusion. Systemic (arterial) blood samples were taken simultaneously with the venous samples at one hour of reperfusion. At the end of the experiment the muscles were harvested and amount of necrosis quantitated by serial transections, nitroblue tetrazolium staining and computerized planimetry. Muscle necrosis on the experimental side was found to be 48.86 +/- 5.37%. Sera were analyzed for TNF activity using a bioassay. TNF levels in the gracilis venous effluent at one hour of reperfusion were not significantly different from the simultaneous systemic (arterial) levels (27.15 +/- 5.05 pg/ml vs 18.23 +/- 4.27 pg/ml). Pre-ischemic levels of TNF were 96.50 +/- 20.12 pg/ml, which was significantly higher than either venous or arterial levels obtained after one hour of reperfusion (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Interleukin-1 and thromboxane release after skeletal muscle ischemia and reperfusion.

Interleukin-1 and thromboxane are known to mediate the host response to sepsis, trauma, and myocardial ischemia. A well-established model of canine isolated gracilis muscle was used to evaluate whether cytokine (interleukin-1) played a role in skeletal muscle ischemia-reperfusion injury. Six adult mongrel dogs (25-30 kg) were subjected to six hours of muscle ischemia followed by reperfusion. Gracilis venous samples were collected pre-ischemia and at one hour of reperfusion. Systemic (arterial) blood samples were taken at one hour of reperfusion. Sera were analyzed for interleukin-1 by bioassay and thromboxane (B2) by radio-immunoassay. The gracilis muscle of the operated limb was harvested in all the animals for assessment of the percentage of muscle necrosis. This was found to be 56.2 +/- 14.8% by serial transections, nitroblue tetrazolium staining, and computerized planimetry. Interleukin-1 levels in the gracilis venous effluent increased from 21.88 +/- 7.13 units/ml during pre-ischemic baseline to 50.42 +/- 9.12 units/ml after six hours of ischemia followed by one hour of reperfusion (p less than 0.04). Thromboxane B2 levels were 2983 +/- 1083 pg/ml and 9483 +/- 2218 pg/ml at pre-ischemia and at one hour of reperfusion respectively (p less than 0.04). Systemic levels of both interleukin-1 and thromboxane B2 at one hour of reperfusion were 0 units/ml and 1584 +/- 520 pg/ml respectively, which were significantly lower than the one hour reperfusion gracilis venous effluent levels (p less than 0.04). This is the first report in which cytokines have been implicated in skeletal muscle ischemia-reperfusion injury.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential susceptibility of inbred mouse strains forecast by acute colonic proliferative response to methylazoxymethanol.

The proliferative response of colonic epithelial cells to methylazoxymethanol (MAM) was followed in 1,2-dimethylhydrazine (DMH)-sensitive SWR/J, moderately resistant C57BL/6J, and resistant AKR/J strains. Untreated AKR mice had a significantly lower labeling index (L1) a shorter proliferative compartment (PC), and a smaller percentage of DNA synthesizing cells in the middle third of the crypts than did the SWR strain. SWR had the highest Ll, the widest PC, and the largest percentage of DNA synthesizing cells in the middle third of the crypts. C57BL/6 mice had characteristics that lay between the sensitive and resistant strains. Pooled data from 1 week after the fifth and sixth injections and 12 weeks after the first MAM injection revealed that extension of the PC had occurred in all strains, but it was only the DMH-sensitive SWR strain that showed extension of the PC to the upper third of the crypt, the greatest shift of proliferating cells to the middle and upper third of crypts, and the greatest increase in Ll. The AKR strain demonstrated these proliferative alterations least, and the C57BL/6 strain fell between them. This differential response to MAM among the strains mimicked that previously reported by us when DMH was investigated. The similarity in response of the colonic epithelial cells of each strain to either a direct-acting (MAM) or indirect-acting carcinogen (DMH) would support the concept that susceptability to this family of carcinogens is directly related to the genetically controlled indigenous proliferative characteristics of the colon.