In many studies using mice, investigators must determine pups' gender at a very early postnatal stage. The gender of mouse pups is typically assessed by measuring the anogenital distance, which is greater in males than in females. This method, however, has proven to be difficult and not completely reliable. The authors describe a quick, easy and reliable method to establish the gender of pigmented mice. In male mice, a pigment spot on the scrotum is visible to the naked eye from the first day of life onwards, whereas female pups lack visible pigmentation in the anogenital region. In lightly pigmented or albino mice, the pigmentation is not obvious or not at all visible. The authors show that identifying this pigment spot is a more accurate and efficient method of determining pup gender compared with measurement of the anogenital distance. This 'spot on' method would therefore be a useful adjunct to conventional methods for determining the gender of pigmented neonatal mice.
Self-reactive T cells have shown to have a potential role as regulators of the immune system preventing or even suppressing autoimmunity. One of the most abundant proteins that can be eluted from human HLA molecules is heat shock protein 70 (HSP70). The aims of the current study are to identify HSP70 epitopes based on published HLA elution studies and to investigate whether T cells from healthy individuals may respond to such self-epitopes. A literature search and subsequent in silico binding prediction based on theoretical MHC binding motifs resulted in the identification of seven HSP70 epitopes. PBMCs of healthy controls proliferated after incubation with two of the seven peptides (H167 and H290). Furthermore H161, H290, and H443 induced CD69 expression or production of cytokines IFNγ or TNFα in healthy controls. The identification of these naturally presented epitopes and the response they elicit in the normal immune system make them potential candidates to study during inflammatory conditions as well as in autoimmune diseases.
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