THE RECEPTOR FOR COMPLEMENT ANAPHYLATOXIN C3A IS EXPRESSED BY MYELOIDCELLS AND NONMYELOID CELLS IN INFLAMED HUMAN CENTRAL-NERVOUS-SYSTEM -ANALYSIS IN MULTIPLE-SCLEROSIS AND BACTERIAL-MENINGITIS

The complement anaphylatoxins C5a and C3a are released at the inflamma tory site, where they contribute to the recruitment and activation of leukocytes and the activation of resident cells, The distribution of t he receptor for C5a (C5aR) has been well studied; however; the recepto r for C3a (C3aR) has only recently been cloned, and its distribution i s uncharacterized, Using a specific affinity-purified anti-C3aR peptid e Ab and oligonucleotides for reverse transcriptase-PCR analysis, C3aR expression was characterized in vitro on myeloid and nonmyeloid cells and in vivo in the brain, C3aR was expressed by adult astrocytes, ast rocyte cell lines, monocyte lines THP1 and U937, neutrophils, and mono cytes, but not by K562 or Ramos, C3aR staining was confirmed by how cy tometry, confocal imaging, and electron microscopy analysis, A 65-kDa protein was immunoprecipitated by the anti-C3aR from astrocyte and mon ocyte cell lysates, Our results at the protein level were confirmed at the mRNA level, Using reverse transcriptase-PCR, Southern blot, and s equencing we found that C3aR mRNA was expressed by fetal astrocytes, a strocyte cell lines, and THP1, but not by K562 or Ramos, The astrocyte C3aR cDNA was identical with the reported C3aR cDNA. C3aR expression was not detected in normal brain sections, However, a strong C3aR stai ning was evident in areas of inflammation in multiple sclerosis and ba cterial meningitis, In meningitis, C3aR was abundantly expressed by re active astrocytes, microglia, and infiltrating cells (macrophages and neutrophils), In multiple sclerosis, infiltrating lymphocytes did not express C3aR, but a strong staining was detected on smooth muscle cell s (pericytes) surrounding blood vessels.