Proteinase-dependent false positive precipitin tests
1972; Elsevier BV; Volume: 49; Issue: 1 Linguagem: Inglês
10.1016/0091-6749(72)90124-8
ISSN1097-6825
AutoresVictoria Wicher, Jerry Dolovich,
Tópico(s)Animal health and immunology
ResumoA variety of nonimmunologic precipitin-like r‘eartions have been descrilwd.l-G In double-diffusion tests in gel, lines rwcmbling precipitin reactions ha\-c been observed between B. szlbfilis alkaline proteinase preparations and the scra of normal individuals with no irltlustrial csposu~ to R. mhtilis products.‘~‘” 0bscrvations on the characteristics of thcsc lines arc dcscrihed here. In double-diffusion tests, the lines tcntlcd to form mow slowly than usual ant&a-antibody precipitates, and they were generall~~ less distinct in appearxnce. They appeared to 1~ washed from the agar during staining with a.mitlo black in an acetic acid-methanol solxnt. There was no relationship betwon these lines and the prcscncc or alwncc of scrwn antibody to the B. suhtilis preparations as determined by ‘9-l aheled antigen-binding methods.11 JVith yariations in cnzpmc i‘anti::cn” eonccntrxtions, lines were elicitctl with all whole normal hunan sera (X118) and with preparations of the third protein I)cali of’ Scl)hadcx C:-200 gel filtration fracl ions (peak 111) of ~IORXI~ IIIUII:II~ Scra, which contains albumin ant1 a,-ant itq-psin. With the corresponding nraci~oriiolcculc-contairiin~ peak I, thcrc wet2 lines which were less dense ant1 loss constantly demonstrated. No such lines were ohserved with IgG-containing peak 11. The lines could be elicited with high concentrations of nonpurified R. subtilis p’otc~inasc preparations and witlt crystalline preparations of the alkaline prot,cinasc, subtilopeptidasc A (SPA). 13. subti7i.v alkaline proteinases arc inactivated 1)~ the selective reaction of nonrevcrsiblc inhibitors such as phcnylnrethancs~~lIhyI fluoride (PXSE’) with the scrinc of the active site of the cr~~yriic.‘~ When pr*otcbirrastl was treated with PBISF, thcw was no apparent loss of antigcnic watt ix-it,y in clouble-cliffnsiori tests with spf‘cific rabbit antisera, but the pryc*il)it in-like reactions with KITS wcrc eliminated. It is therefore concluded that that i’ormation of the lines is dcpcntlent upon the proteinase activity of the ‘iantigen” preparations. 1 ,ines could be readily produced in I$$ hours by coanterirnnrurroelectrol~lro~(~sis iC1 HI’) l3 (Fig. I). With this tcchniquc. slcntLcr lines of low density could
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