Comparison between enzyme-linked immunosorbent assay and cytotoxic cross-match procedures for detecting IgG anti-donor antibodies

BACKGROUND: Disadvantages inherent to complement-dependent cytotoxicity cross-match (CDC XM) methods are the requirements for complement and viable target cells, detection of antibodies (Abs) against non-HLA antigens, and subjective scoring. Cross-Stat (SangStat Medical Corp., Menlo Park, CA), a recently developed enzyme-linked immunosorbent assay XM procedure for the detection of IgG anti-donor HLA Abs, is theoretically devoid of these flaws. METHODS: We compared results of Cross-Stat and our standard anti-human globulin (AHG)-enhanced CDC XM procedure on 524 sera from 230 transplant candidates, which were evaluated against 51 cadaveric donors. RESULTS: There was a significant correlation between AHG-CDC IgG XM and Cross-Stat results (P<0.001). For false negative sera, repeat AHG-CDC IgG XMs were still positive after platelet absorption, indicating that the Abs present were either non-HLA Abs or anti-HLA class II. Flow cytometry testing of false positive sera usually (42/62) substantiated Cross-Stat results, indicating that the discrepancy with AHG-CDC IgG XM is caused by greater sensitivity of Cross-Stat. Relative to the AHG-CDC XM, the sensitivity of Cross-Stat was 100%, the specificity was 93%, the positive predictive value was 73%, and the negative predictive value was 100%. A technical shortcoming of the Cross-Stat assay is that the frequency of indeterminate samples in the assays was 15%. Among 49 Cross-Stat negative vs. 13 Cross-Stat positive primary cadaveric renal allograft recipients (all AHG-CDC IgG-XM negative), there was no statistical difference in overall graft survival. CONCLUSION: Given the important theoretical advantages of enzyme-linked immunosorbent assay-based XM methods over the CDC XM, however, further testing of the clinical relevance of the Cross-Stat is warranted.     

Pattern formation in chick feather development: distribution of beta 1-integrin in normal and scaleless embryos

We have examined the immunolocalization of beta 1-integrin during feather development in the spino-lumbar tract of the backskin from normal and scaleless chick embryos. beta 1-integrin appears during early feather development in three distinct phases which correspond to important developmental events. The first phase (5-5 1/2 days of incubation; Hamburger and Hamilton [H.H.] stage 27) represents the period prior to the formation of dermis. During this phase, beta 1-integrin antiserum labels mesenchymal cells located in the central region of the spino-lumbar tract where the initiation site for feather development is located. The second phase (5 1/2-7 1/2 days of incubation; H.H. stages 28-32) corresponds to the period during which dermis is formed. The cells that make up the dermis are readily distinguished by their lack of beta 1-integrin immunostaining. The third phase (7 1/2-10 days of incubation; H.H. stages 33-36) begins with the sudden appearance of beta 1-integrin in the central and lateral regions of the dermis. The pattern of beta 1-integrin immunostaining in scaleless backskin becomes different from that of normal backskin during this phase. In normal backskin the dermal condensations of feather germs are not labeled with the beta 1-integrin antiserum. This produces a heterogeneous immunostaining pattern very similar to the pattern seen for Type I collagen (Mauger et al. [1982] Dev. Biol. 94:93-105). In contrast, homogeneous immunostaining is observed in the dermis of scaleless backskin. The initial time of appearance, manner of appearance, and pattern of integrin expression in the third phase suggest that beta 1-integrin may be involved in the stabilization of the feather pattern. We also observed the appearance of beta 1-integrin on the epidermal basal cells during the time of feather follicle formation. The beta 1-integrin antiserum reacts strongly with the baso-lateral surfaces of normal basal cells, yet the basal surfaces of the scaleless basal cells are unstained. This lack of immunostaining along the basal surfaces of the scaleless basal cells may relate to the abnormal adhesion between the epidermis and dermis in scaleless backskin.     

Microcomputer compilation and solution of crosswords

A system is described which enables crosswords to be automatically compiled or solved, or which alternatively may assist in their manual compilation or solution. Illustrative examples are quoted and the performance of the system, which has been written for an Acorn BBC model B microcomputer with disc pack and printer, is discussed.