Effects of high and low molecular soluble factors of the bone marrow on antibody formation and pain sensitivity in animals

In addition to the immunostimulating activity, bone marrow mediators, myelopeptides (MP) show the dose-dependent effect on the development of pain sensitivity in mice. When injected in nanogram amounts, MP induce hyperalgesia and 3-9 fold higher production of antibodies against SREC. When injected in milligram amounts, they exhibit hypoalgesic effect and no influence on antibody production. Immunostimulating effect in MP (mol, mass less than 1 KD, fraction 3) is accompanied with hypoalgesia. Bone marrow factors of mol. masses 40-150 KD (fraction 1) eluted at Sephadex G-25 gel-filtration before MP enhance the pain sensitivity tHreshold and show a potent immunodepressive effect. Thus the bone marrow factors are capable of exhibiting the opposite effects on the immune system in the pain control system that evidences the tight interrelation between these systems.     

High levels of expression of full length human pro-alpha 2(V) collagen cDNA in pro-alpha 2(V)-deficient hamster cells

A full length cDNA encoding human pro-alpha 2(V) collagen was constructed. Partial sequencing of the cDNA and primer extension analysis of mRNA from fibroblasts found that pro-alpha 2(V) mRNA differs from the mRNAs of other fibrillar collagens in the increased length of its 5'-untranslated region. The pro-alpha 2(V) cDNA was placed downstream of the human cytomegalovirus immediate early promoter/regulatory sequences for expression studies in cultured Chinese hamster lung cells. These cells have been shown previously to synthesize large quantities of pro-alpha 1(V) homotrimers as their only collagenous product. Transfection resulted in a number of clonal cell lines that express human alpha 2(V) RNA at levels comparable to, and in some cases greater than, levels found in normal human skin fibroblasts. Pro-alpha 2(V) chains produced in the majority of clonal lines were of sufficient quantity to complex all available endogenous pro-alpha 1(V) chains. Chimeric heterotrimers, composed of hamster alpha 1(V) and human alpha 2(V) chains in a 2:1 ratio, were stable to pepsin digestion and were found predominantly associated with the cell layer. Surprisingly, pro-alpha 2(V) chains, in excess to pro-alpha 1(V) chains, were found in the extracellular matrix and, in much greater abundance, in media. These chains were pepsin sensitive, indicating that pro-alpha 2(V) chains can be secreted as nonstable homotrimers or as free chains.