Garratty G, Petz L D
Transfusion. 1976 Jul-Aug;16(4):297-306. doi: 10.1046/j.1537-2995.1976.16476247049.x.
Human red blood cells, sensitized with complement in vivo and by a variety of methods in vitro, (e.g. blood group antibody, low ionic strength, alternate pathway), were tested with a battery of anti-complement sera (anti-C3, -C3c, -C3d, -C4, -C4c). Red blood cells could be prepared by relatively simple methods to yield cells sensitized with C3 and C4, C3 but not C4, C4 but no C3, C3d with no C3c and C4d with no C4c. These cells are suitable for standarization and quality assurance of antiglobulin sera (AGS). Anti-C3d is necessary for optimal detection of sensitization of red blood cells by complement in vivo by the direct anti-globulin test (DAT). Anti-C3d may also be optimal for the indirect antiglobulin test (IAT) especially if incubation periods greater than one hour are employed. Potent anti-C4 and anti-C3 antisera made in the authors' laboratory resulted in numerous weakly positive antiglobulin tests when testing red blood cells from refrigerated clots (especially anti-C4) but red blood cells from refrigerated anticoagulated segments gave negative results. When red blood cells were incubated in normal serum at room temperature (as in the room temperature phase of a compatibility test), some positive results were again obtained with the potent anti-C4 and anti-C3 antisera. However, one commercial antiglobulin serum containing anti-complement antibodies that were at least as potent as any other commercial antiglobulin serum gave uniformly negative results under the above conditions. Anti-C4 antibodies may be omitted from anti-globulin sera without decreasing the efficacy of such antisera to be used in compatibility testing. Thus, positive results in the compatibility test due to detection of clinically insignificant cold antibodies in the IAT by the anti-complement antibodies in AGS, may be avoided if anti-C4 is omitted or is in low concentration and if the concentration of anti-C3d is carefully standardized. A higher concentration of anti-C3d could be used for compatibility tests if red blood cells from anticoagulated segments were used instead of those from clots and if a separate tube were used for the IAT at 37 C rather than using one tube for both room temperature and 37 C incubations.
用人源红细胞进行实验，这些红细胞在体内以及通过多种体外方法（例如血型抗体、低离子强度、替代途径）致敏补体后，用一系列抗补体血清（抗C3、抗C3c、抗C3d、抗C4、抗C4c）进行检测。通过相对简单的方法可以制备红细胞，使其分别被C3和C4致敏、仅被C3而非C4致敏、仅被C4而非C3致敏、仅被C3d而非C3c致敏以及仅被C4d而非C4c致敏。这些细胞适用于抗球蛋白血清（AGS）的标准化和质量保证。抗C3d对于通过直接抗球蛋白试验（DAT）最佳检测体内补体致敏的红细胞是必需的。抗C3d对于间接抗球蛋白试验（IAT）可能也是最佳的，特别是在孵育时间超过一小时的情况下。作者实验室制备的高效抗C4和抗C3抗血清，在检测来自冷藏血块的红细胞（尤其是抗C4）时，导致大量弱阳性抗球蛋白试验结果，但来自冷藏抗凝段的红细胞检测结果为阴性。当红细胞在室温下于正常血清中孵育时（如在相容性试验的室温阶段），高效抗C4和抗C3抗血清再次得到一些阳性结果。然而，一种含有抗补体抗体且效力至少与其他任何市售抗球蛋白血清相当的市售抗球蛋白血清，在上述条件下给出的均为阴性结果。抗球蛋白血清中可以省略抗C4抗体，而不会降低此类抗血清在相容性试验中的效力。因此，如果省略抗C4或其浓度较低，并且抗C3d的浓度经过仔细标准化，那么可以避免由于AGS中的抗补体抗体在IAT中检测到临床上无意义的冷抗体而导致的相容性试验阳性结果。如果使用来自抗凝段的红细胞而非来自血块的红细胞，并且在37℃下为IAT使用单独的试管，而不是在室温及37℃孵育都使用同一试管，那么可以使用更高浓度的抗C3d进行相容性试验。
