Oxford: Oxford University Press; 1995. 1% Nonidet P-40 (Sigma, St Louis, MO, USA) in 110 mM NaCl and 50 mM Tris (pH 75) in the presence of 50 were purified by affinity chromatography with lentil lectin-sepharose (Amersham Pharmacia Biotech Gemcabene calcium Abdominal) and anti-Fcin plasma from any donors is possible, we selected NA(1 +, 2-) phenotyped donors to compare the levels of sFchealthy settings * 001) Gemcabene calcium or OA individuals ? 001). Three hundred and forty-two healthy volunteers were recruited randomly from the hospital staff. One hundred individuals were selected for an NA(1 +, 2-) phenotype to constitute the pooled plasma. Forty-one age-matched individuals were selected as healthy settings. None of the control individuals had any evidence of renal, hepatic, infectious or inflammatory disease or diabetes mellitus, and none were taking any medication. FcRIIIB-NA(1, 2) genotyping assays Genomic DNA (gDNA) was extracted from leucocytes by standard techniques. Genotyping for the Fcconcentrations were measured with immuno-PCR relating to Furuya in the plasma pool was arranged at 100 AU. to pooled plasma, measured sFcwas prepared from tradition Gemcabene calcium supernatant of monocytes by affinity chromatography with Lentil Lectin-Sepharose and anti-FcRIII MoAb CLBFcRgranI-Sepharose, as explained before. Contaminating MoAb p105 was eliminated by filtration having a 100-kDa cut-off membrane (OMEGA? disk, Pall Filtron Co., Northborough, MA, USA), followed by concentration of Fc(OD280: 0457). Purified sFcor PBS (1/10 volume) was supplemented to pooled plasma from healthy NA(1 +, 2-) phenotyped donors and then sFcwas added to the pooled plasma or to the RA patient’s plasma, and consequently the amount of the three types of sFctest. Correlations were tested by Bartlett’s test and multiple comparisons by analysis of variance. RESULTS FcRIIIaM-specific anti-FcRIII MoAb, MKGR14 We developed a new anti-Fcfrom the lysate of cultured monocytes, but did not precipitate Fcin plasma. As demonstrated in Fig. 3, the level of sFcin RA individuals was about four instances higher than that in healthy settings. In particular, there were extraordinarily higher levels in individuals in stage III or IV. In contrast, the Gemcabene calcium level of total sFc(top), sFc 005, ** 001), OA individuals (# 005, ## 001) or stage 4 ( 005, 001). As demonstrated in Table 3, the sFclevels in plasma correlated with age, the sFcto sFc= 0335**, sFcto total sFc= 0274*, sFc= 0447**) in RA individuals, but not in healthy settings. In RA individuals, there was a significant correlation between the sFclevels and the Lansbury Index, and between the sFc(remaining), sFcindicates level of sFcto pooled plasma, measured sFcwere 26378 961 AU in sFcassay, 9578 738 AU in sFcprepared from tradition supernatant of monocytes and the should be developed. We have succeeded in raising a new anti-Fcpresent was about half that of sFcand the varied between RA individuals. In RA individuals, both the sFclevels correlated with the Lansbury Index (Fig. 4, Table 3) and thus with the disease severity. However, there was a difference between the two levels; i.e. sFclevels were increased directly proportional to C-reactive protein (Table 3) and thus to inflammation. sFccorrelated individually with the severity of RA. In conclusion, sFcmay serve as a new marker of the disease activity in RA. Acknowledgments We say thanks to Dr Masja de Haas and Dr Federico Garrido for his or her generous gifts of antibodies and Mr Takashi Kizu, Mr Hideaki Nakatuji and Mr Hiroki Bandoh for his or her assistance with the genotyping assay. This work was supported partly by grants from your Ministry of Education, Grant-in-Aid for Scientific Study (C) (nos 07672505 and 10672192), from your Charitable Trust Clinical Pathology Study Basis of Japan and from your TERUMO Life Technology Basis (no. 99C319). Referrals 1. Ravetch JV, Perussia BV. Alternate membrane forms of Fcculture conditions. Am J Resp Cell Mol Biol. 1991;5:307C14. [PubMed] [Google Scholar] 5. Huizinga TW, de Haas M, vehicle Oers MH, et al. The plasma concentration of soluble Fcreceptors (sFcand receptor type III (CD16) Eur J Immunol. 1998;28:2101C7. [PubMed] [Google Scholar] 17. Koene HR, de Haas M, Kleijer M, et al. NA-phenotype-dependent variations in neutrophil Fc em /em RIIIb manifestation cause Gemcabene calcium variations in plasma levels of soluble Fc em /em RIII. Br J Haematol. 1996;93:235C41. [PubMed] [Google Scholar] 18. de Haas M, Kleijer M, von Roos D, von dem Borne AEGKr. Characterization of MAbs of the CD16 cluster and six fresh generation CD16 MAbs. In: Schlossmann SF, et al., editors. Leukocyte typing V. Oxford: Oxford University or college.
