Human melanoma cells inhibit the earliest differentiation steps of human Langerhans cell precursors but failed to affect the functional maturation of epidermal Langerhans cells

Tumour-derived factors suppress differentiation and function of in vitro generated DC. Here, we investigate the effect of two melanoma clones differing in their invasive and metastatic properties on the generation and/or functional maturation of human epidermal LC. LC were generated from CD34+ cord blood progenitors under GM-CSF/TNF-α/TGF-β1. CD34+ cells were co-cultured with or without melanoma cells using Transwell dishes. After 11 days of co-culture, CD34+-derived cells display a non-adherent undifferentiated morphology, a high level of monocytic CD14 marker, a down-regulated expression of LC markers (CD1a, E-cadherin) and DC markers (CD40, CD80, CD54, CD58, CD83, CD86, HLA-DR, HLA-class I). These cells were less potent than control LC in inducing allogeneic T cell proliferation. The generation of the CD14+ population was correlated with a decrease in the CD1a+ population, without any statistical differences between the two clones. Melanoma cells diverted the differentiation of CD34+ cells towards a dominant CD14+ population only if the progenitors were in an early growth phase. IL-10, TGF-β1 and VEGF were not responsible for these effects, as assessed by using blocking antibodies. By contrast, co-culture of fresh epidermal LC with melanoma cells did not affect their phenotype and function. Our data demonstrate that melanoma cells inhibit the earliest steps of LC differentiation, but failed to affect the functional maturation of epidermal LC. This suggests that melanoma cells participate in their own escape from immunosurveillance by preventing LC generation in the local cutaneous microenvironment. © 2001 Cancer Research Campaign http://www.bjcancer.co

Cut site preference allows influenza A virus PA-X to discriminate between host and viral mRNAs

Many viruses block host gene expression to take over the infected cell. This process, termed host shutoff, is thought to promote viral replication by preventing antiviral responses and redirecting cellular resources to viral processes. Several viruses from divergent families accomplish host shutoff through RNA degradation by endoribonucleases. However, viruses also need to ensure expression of their own genes. The influenza A virus endoribonuclease PA-X solves this problem by sparing viral mRNAs and some host RNAs necessary for viral replication. To understand how PA-X distinguishes between RNAs, we characterized PA-X cut sites transcriptome-wide using 5' rapid amplification of complementary DNA ends coupled to high-throughput sequencing. This analysis, along with RNA structure predictions and validation experiments using reporters, shows that PA-Xs from multiple influenza strains preferentially cleave RNAs at GCUG tetramers in hairpin loops. Importantly, GCUG tetramers are enriched in the human but not the influenza transcriptome. Moreover, optimal PA-X cut sites inserted in the influenza A virus genome are quickly selected against during viral replication in cells. This finding suggests that PA-X evolved these cleavage characteristics to preferentially target host over viral mRNAs in a manner reminiscent of cellular self versus non-self discrimination

Phylogenetic Studies of the Bullous Pemphigoid Antigen-1 Using Human Monoclonal-antibodies

Bullous pemphigoid is an autoimmune skin disorder with production of autoantibodies against bullous pemphigoid antigen 1 (BPAg1) and bullous pemphigoid antigen 2 (BPAg2) which are constitutively expressed in hemdesmosomes. Phylogenetic study of the reactivity of 3 human monoclonal antibodies (HuMabs) specific for BPAg1 was performed using immunohistochemical analysis of skin sections. The serum of the 2 BP patients from which the 3 HuMabs were derived stained the basement membrane zone on skin cryostat section of all species tested including fishes, amphibians, reptiles, birds and mammals. However, of the 3 HuMabs BP1, BP2 and BP3, only BP3 reacted with the skin of reptiles, birds and mammals while Bp1 and Bp2 exclusively bound to mammalian skin. These data provide evidence for the existence of multiple epitopes on BPAg1 able to bind autoantibodies

IgM autoantibodies to 180- and 230- to 240-kd human epidermal proteins in pregnancy

BACKGROUND AND DESIGN: A previous study has suggested that there is a novel entity among the polymorphous eruptions of pregnancy (PEP) associated with circulating anti-basement membrane zone IgM autoantibodies. To determine if the presence of anti-basement membrane zone IgM autoantibodies is a feature of PEP, serum samples from 52 patients with a PEP, 69 healthy pregnant women, and 42 nonpregnant women were prospectively evaluated by indirect immunofluorescence using salt-split human skin as substrate. Serum samples were also tested by immunoblotting using keratinocyte extracts and anti-human IgM antibodies. The reactivity of some serum samples was examined using two recombinant bullous pemphigoid antigen proteins. RESULTS: The percentage of women with a PEP, healthy pregnant women, and nonpregnant women who had anti-basement membrane zone IgM antibodies by indirect immunofluorescence was similar: 12%, 10%, and 14% of cases, respectively. By immunoblotting, 14% of the serum samples from the patients with a PEP, 12% of the serum samples from the healthy pregnant women, but only 2% of the serum samples from the nonpregnant women contained IgM antibodies that reacted with epidermal proteins of 180 and/or 230 to 240 kd. The recombinant bullous pemphigoid antigen proteins were not recognized by any of the serum samples that showed a reactivity by immunoblotting using keratinocyte extracts. CONCLUSION: There is no evidence for the existence of a novel entity of pregnancy defined by circulating anti-basement membrane zone IgM autoantibodies. Immunoblotting detects IgM autoantibodies that react with epidermal proteins of 180 and/or 230 to 240 kd. These autoantibodies appear to be more frequent in pregnant than in nonpregnant women. Although the nature of the target antigen(s) remains to be established, pregnancy may be associated with low levels of IgM autoreactivity against epidermal proteins

Prenatal diagnosis of cerebellar cortical dysplasia associated with abnormalities of foliation.

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