The objectives of the Fifth International BoLA Workshop were to: standardize nomenclature, compare typing methods, and characterize BoLA haplotypes. The workshop was based on the distribution of blood samples (cells) from 60 selected cattle to 14 laboratories. Results for the class I (BoLA-A) region are presented in this paper while results for the class II regions are presented in a separate report. Thirty-six of the 50 previously established serological class I specificities were represented in the cell panel. However, only 30 specificities could be confirmed. Two specificities, A16 and A32, were upgraded from provisional, workshop (w) specificities to BoLA-A locus specificities and three new specificities, w51(w28), w52 and w53(w28), were defined. The 39 specificities distinguished 30 class I haplotypes in the 60 animals. Class I isoelectric focusing proved to be a useful adjunct to the serology. Isoelectric focusing confirmed several serologically defined splits and detected splits of A15(A8), A18(A6) and A22(w49) that had not been detected by serology. Subsequently, serological support for splits of A15(A8) and A22(w49) was found.

Adhesion molecule expression was analysed on porcine blood and lymphoid organ CD4+ CD8 naive T helper (Th) lymphocytes, CD4+CD8+ memory Th lymphocytes (particular to the pig), CD4-CD8high cytotoxic T (Tc) lymphocytes, CD4 CD8low NK cells (CD3- in the pig), CD4-CD8- T-cell receptor-gammadelta-positive (TCRgammadelta+) lymphocytes, B lymphocytes and monocytes. While CD44 expression was relatively homogeneous amongst mononuclear cells, differences were noted for the integrins. Blood naive Th lymphocytes were CD49d(low)CD11a(low), as were splenic naive Th cells; blood memory Th lymphocytes were CD49d(high)CD11a(low), splenic memory Th cells were CD49d(high)CD11a(high) with a CD49d(high)CD11a(low) subpopulation; blood Tc lymphocytes were mainly CD49d(low)CD11a(low), and splenic cells were CD49d(high) CD11a(high). Lymph node lymphocytes were more homogeneous in their integrin expression. These were relatively CD49d(low)CD11a(low), except the memory Th lymphocytes which had higher integrin expression. B lymphocytes related to the majority of integrin(low) T cells, while monocytes and NK cells were CD49d(high) CD11a(high); gammadelta T lymphocytes showed variable CD49d expression but a CD11a(high) phenotype. CD49d(high) CD11a(high) co-expression was found, and this phenotype was typical of, but not exclusive to, CD25+ (activated) lymphocytes. These results demonstrated that porcine memory Th lymphocytes and NK cells, as well as activated cells, would have increased integrin-dependent activities compared with naive Th lymphocytes, and integrin-dependent reactions would probably vary between blood and lymphoid organ cells.

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The incidence of subclinical mastitis in Simmental and Simmental-Red Holstein cattle in relation to the bovine major histocompatibility complex (BoLA) was investigated. Quarter milk samples of 166 cows consisting of fifteen halfsib groups of different ages and lactation stages were analysed for somatic cell counts (SCC) and bacteriological infection. From these data the udder health status (UHS) of the animal was determined. Each cow was typed serologically for BoLA class I and class II specificities. The statistical evaluation for UHS was performed using a logistic regression model. The effect of BoLA on SCC was estimated by least square analysis. Animals carrying the BoLA class II haplotype "b" were significantly more affected by subclinical mastitis than those without this haplotype. Breed group showed a significant influence on both UHS and corrected mean logSCC (P < 0.0001 resp. P < 0.05). Lactation stage had a significant effect on SCC but only a weak influence on UHS (P < 0.0001 resp. P < 0.13). ZUSAMMENFASSUNG: Mögliche Assoziation zwischen dem bovinen Haupthistokompatibilitätskomplex und subklinischer Mastitis In dieser Studie wurde die Beziehung zwischen dem bovinen Histokompatibilitätskomplex (BoLA) und der Prävalenz subklinischer Mastitis bei Simmentaler und mit Red Holstein eingekreuzten Simmentaler Kühen untersucht. Bei 166 Kühen, aus 15 Halbgeschwistergruppen, unterschiedlichen Alters und in verschiedenen Laktationsstadien, wurden Milchproben entnommen und auf Zellzahl (SCC) und bakterielle Infektion untersucht. Aus diesen Daten wurde ein Eutergesundheitsstatus (UHS) definiert. Jede Kuh wurde serologisch für BoLA Klasse I und Klasse II Spezifitäten typisiert. Die statistische Auswertung für den UHS erfolgte mit einem logistischen Regressionsmodel. Der Einfluß von BoLA-Haplotypen auf SCC wurde mit der Least Square Analyse ermittelt. Tiere mit dem Klasse II Allel "b" zeigten mehr Euterprobleme als Kühe ohne dieses Allel. Die Rassegruppe übte sowohl auf den UHS wie auch auf den korrigierten Mittelwert der Zellzahlen einen signifikanten Einfluß aus (P < 0.0001 resp. P < 0.05). Der Effekt des Laktationsstadiums auf die Zellzahl war signifikant, aber für den UHS wurde nur ein schwacher Einfluß des Laktationsstadiums festgestellt (P < 0.0001 resp. P < 0.13).

Blood samples from 54 animals were exchanged between 15 laboratories in nine countries to improve and expand BoLA class 1 and class I1 typing. A total of 27 out of 33 (82%) of previously accepted BOLA-w specificities were represented within the cell panel. Seventeen new serum-defined BoLA specificities were accepted by the workshop participants, thus expanding the number of internationally recognized BoLA specificities to 50. The large number of new specificities detected resulted from the number of serological reagents used ( n = 1139) and the genetic diversity of the cell panel. Confidence derived from the high percentage of agreement between the laboratories on antigen detection (97.3%; r = 0.84) permitted the removal of the workshop (w) notation from 23 BOLA-w specificities and their acceptance as full status BOLA-A antigens. Two new non-BOLA antigens were also detected, one completely included within the red blood cell factor S' (BoLy-S'), whereas a second (BoLy-wl) did not show any association with tested red blood cell factors. A comparison between serological, isoelectric focusing (IEF) and DNA typing for BoLA class I1 polymorphism was conducted with a subset of workshop cells. Correlation between the three methods was significant for three combinations of alleles. Three other serologically defined class I1 specificities were correlated with DR and/or DQ restriction fragment length polymorphism (RFLP) types, whereas six additional IEF types were correlated with DR and/or DQ RFLP types ( r L 0.50). Several new IEF, DRB, DQA and DQB RFLP patterns were identifed. In 46 animals that were typed for BOLA-DR and DQ genes by RFLP analysis, 46 different BoLA haplotypes were tentatively defined. These 46 haplotypes were distinguished by 31 serologically-defined BOLA-A alleles (and 2 'blanks'), 15 DRB RFLP types (plus up to 10 new DRB RFLP patterns) and 23 DQA-DQB haplotypes.