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Immunohematology JOURNAL of BLOOD GROUP SEROLOGY and EDUCATION Immunohematology JOURNAL OF BLOOD GROUP SEROLOGY AND EDUCATION VOLUME 18, NUMBER 1, 2002 From the publishers of Immunohematology A Comprehensive Laboratory Manual Immunohematology Methods and Procedures Featuring— •Over 100 methods— just about every method used in a reference lab. • Eleven chapters discussing problems faced by blood group serologists and the procedures and methods that can be used to solve them. • An extra set of the methods to use at the bench, printed on durable waterproof paper. • See business reply order card enclosed in this issue or order on the Web at redcross.org/immunohematology Immunohematology JOURNAL OF BLOOD GROUP SEROLOGY AND EDUCATION VOLUME 18, NUMBER 1, 2002 CONTENTS 1 A review of the Knops blood group: separating fact from fallacy J.M. MOULDS 9 A gel microtyping system for diagnosis of paroxysmal nocturnal hemoglobinuria B. ZUPANSKA, B. BOGDANIK, AND H. PYL 13 Antibody screening in 37°C saline. Is it safe to omit it using the indirect antiglobulin (gel) test? J. DURAN AND M. FIGUEIREDO 16 Autoanti-D in a patient after cladribine treatment for lymphoplasmocytic lymphoma J. CID, V. BELTRAN, L. ESCODA, E. ELIES, AND C. MARTIN-VEGA 19 Warm autoimmune hemolytic anemia with mimicking anti-c and -E specificities H.-Y. HSIEH, D. MORONEY, D. NAUMANN, J. HATA, N. VOSNIDOU, R. KESSINGER, N. SHAHAB, N. HAKAMI, AND D. SMITH 23 COMMUNICATIONS Letters to the Editors Anti-cE (Rh27), a rarely occurring antibody G. LODI, D. RESCA, R. REVERBERI, AND M. GOVONI September 11 D. LONG Letter From the Editors Thoughts on September 11 24 24 ANNOUNCEMENTS ADVERTISEMENTS 26 INSTRUCTIONS FOR AUTHORS EDITOR-IN-CHIEF MANAGING EDITOR Delores Mallory, MT(ASCP)SBB Mary H. McGinniss, AB, (ASCP)SBB Rockville, Maryland Bethesda, Maryland TECHNICAL EDITOR MEDICAL EDITOR Christine Lomas-Francis, MSc S. Gerald Sandler, MD Austin, Texas Washington, District of Columbia EDITORIAL BOARD Patricia Arndt, MT(ASCP)SBB Brenda J. Grossman, MD Scott Murphy, MD Los Angeles, California St. Louis, Missouri Philadelphia, Pennsylvania James P. AuBuchon, MD Christine Lomas-Francis, MSc Paul M. Ness, MD Lebanon, New Hampshire Austin, Texas Baltimore, Maryland Malcolm L. Beck, FIBMS, MIBiol Gary Moroff, PhD Mark Popovsky, MD Kansas City, Missouri Rockville, Maryland Braintree, Massachusetts Richard Davey, MD Ruth Mougey, MT(ASCP)SBB Marion E. Reid, PhD, FIBMS New York, New York Carrollton, Kentucky New York, New York Geoffrey Daniels, PhD John J. Moulds, MT(ASCP)SBB Susan Rolih, MS, MT(ASCP)SBB Bristol, United Kingdom Raritan, New Jersey Cincinnati, Ohio Sandra Ellisor, MS, MT(ASCP)SBB Marilyn K. Moulds, MT(ASCP)SBB David F. Stroncek, MD Raritan, New Jersey Houston, Texas Bethesda, Maryland George Garratty, PhD, FRCPath Marilyn J. Telen, MD Los Angeles, California Durham, North Carolina PRODUCTION ASSISTANT Linda Berenato COPY EDITOR Lucy Oppenheim ELECTRONIC PUBLISHER James Altes Immunohematology is published quarterly (March, June, September, and December) by the American Red Cross, National Headquarters, Washington, DC 20006. The contents are cited in the EBASE/Excerpta Medica and Elsevier BIOBASE/Current Awareness in Biological Sciences (CABS) databases. The subscription price is $30.00 (U.S.) and $35.00 (foreign) per year. Subscriptions, Change of Address, and Extra Copies: Immunohematology, P.O. Box 40325 Philadelphia, PA 19106 Or call (215) 451-4902 Web site: redcross.org/immunohematology or redcross.org/pubs/immuno Copyright 2002 by The American National Red Cross ISSN 0894-203X A review of the Knops blood group: separating fact from fallacy J.M. MOULDS Introduction blood group system, i.e., International Society of Blood It has been more than 10 years since the topic of “high-titer, low- Transfusion (ISBT) 011. Since that time, all have avidity” (HTLA) antibodies was reviewed in Immunohematology. We have learned a lot about these antibodies in the past 10 years achieved blood group status with the exception of Cost and that knowledge has helped us to understand some of the (Csa), which remains a “collection” as defined by the unusual characteristics of these antibodies. Furthermore, it has ISBT Working Party on Terminology of Red Cell Surface helped us to name and delineate the various associated blood Antigens. Chido and Rodgers were the first antigens in group systems. Although we will begin with a general review of HTLAs, this manuscript will focus on the recent findings in the the group to be located on a membrane protein and Knops blood group system. Immunohematology 2002;18:1–8. given blood group status (ISBT 017). They were found Key Words: Knops, polymorphism, CR1, blood group to be antigens carried on the C4d fragment of the C4B and C4A isotypes, respectively. The Knops antigens were identified as polymorphisms of complement Historical Perspective receptor type one (CR1) and accordingly Knops became the 22nd blood group system. Serologic and HTLAs biochemical studies resulted in the assignment of both To really understand some of the fallacies that have Holley and Gregory to the Dombrock system (ISBT 014) arisen regarding HTLAs, we must delve into their and, most recently, JMH (CDw108) was named system history, i.e., how they were discovered and how they 026.1 were named. In the late 1960s and early 1970s, blood group serologists from the American Association of Knops Blood Banks (AABB) and the American Red Cross (ARC) Although many of the HTLA antibodies were reference laboratories would gather at wet workshops simultaneously being investigated in the 1960s, one of and share samples for the classification of antibodies the first to be reported was anti-Csa. Giles et al.2 found and the formation of blood group systems. During this that the sera from three patients had the same time a number of weak antihuman globulin-reactive specificity and they were mutually compatible. Thus, (AHG) antibodies that appeared to have similar Csa (Cost) was named after two of the first antibody characteristics were studied. When the serum was producers, i.e., Copeland (CO) and Stirling (ST). titrated and the reactions examined microscopically, Although anti-Csa appeared to have some association positive reactions could be observed at dilutions of 1:64 with York, it would later be shown not to be part of the or greater. This was very unusual, as the neat reactions Knops blood group system.3,4 would only be m+ to 1+. The agglutinates often were The Knops system began to take form when anti-Kna fragile and could be easily “shaken out.” Hence, several was described in a transfused Caucasian female who reference laboratory technologists began referring to had a saline-reactive anti-K plus an unidentified this group as “high-titer, low avidity” antibodies. They antiglobulin-reactive antibody to a high-frequency never meant this to denote a blood group system, only antigen.5 A blood bank technologist (Helgeson) found an identifying characteristic (D. Mallory, personal that her red blood cells (RBCs) were compatible with communication). the Knops serum. The “Helgeson phenotype” would Members of this newly formed group included: later be identified as the serologically null phenotype Chido, Rodgers, Knops, McCoy, Swain-Langley, Cost, for the Knops blood group. York (Yka) was the next York, JMH, Holley, Gregory, and, sometimes, Cartwright. high-incidence Knops antigen (KN5) to be reported but The latter was already known to be an independent it was initially believed to be associated with Cost (Csa) IMMUNOHEMATOLOGY, VOLUME 18, NUMBER 1, 2002 1 J.M. MOULDS rather than Kna.6 Several years later, Molthan and that could disrupt disulfide bonds, i.e., dithiothreitol Moulds7 described a new antigen, McCa, that seemed to (DTT) and 2-aminoethylisothiouronium bromide (AET), be related to Kna. Interestingly, a majority of McCa could also destroy Knops, McCoy, and York. This was antibody producers were black, while most of those the dogma, although we had little insight into why making anti-Kna were Caucasian, thus suggesting that these antibodies behaved as they did. ethnic differences might exist in their respective gene frequencies. Finally, Sla and Vil were reported in separate Biochemical Identification of Knops abstracts, with one author using the term McCc for Sla.8,9 Antigens In 1991, two groups identified CR1 as the protein Antigen/Antibody Characteristics carrying the Kna, McCa, and Sla (McCc) blood group The description of HTLA antibody characteristics and antigens.3,10 In addition, Moulds et al.3 identified Yka on their corresponding antigens has remained fairly CR1 and suggested that the Helgeson phenotype was unchanged over the past 30 years. Prominent due to low CR1 copy numbers on the RBCs (E-CR1). characteristics of the Knops system antibodies and The CR1 gene also exhibited two other polymorphisms besides the Knops blood group. A structural poly- Table 1. Prominent characteristics of Knops system antibodies morphism results from four different alleles that encode • High-titer, low-avidity • Can demonstrate variable reactions four different molecular-weight kiloDalton (kD) • Not neutralized by pooled serum or other body fluids proteins: 190 kD (CR1*3), 220 kD (CR1*1), 250 kD • Difficult to adsorb and elute • IgG, reacting by AHG* technique (CR1*2), and 280 kD (CR1*4). The third commonly • Do not bind complement recognized polymorphism is based upon quantitative • Reactive with enzyme-treated RBCs† • Usually not clinically significant differences in E-CR1. A Hind III restriction fragment length polymorphism (RFLP), identified in Caucasians, is
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