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Immunohematology basics


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Basics for all healthcare workers

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Immunohematology basics

  1. 1. Deepa Babin @TMC Kollam
  2. 2. <ul><li>The study of immunologic reactions involving all components of blood </li></ul><ul><li>Deals with antigens, antibodies and antigen-antibody reactions </li></ul>IMMUNOHEMATOLOGY Deepa Babin @TMC Kollam
  3. 3. Application in .... <ul><li>transfusion of blood & its components </li></ul><ul><li>diagnosis, prevention & management of immunization asso with pregnancy </li></ul><ul><li>leukocyte testing for organ transplantation </li></ul><ul><li>laboratory resolution of parentage problems </li></ul>Deepa Babin @TMC Kollam
  4. 4. Agglutination / Hemolysis Deepa Babin @TMC Kollam
  5. 5. Complete Antibodies Deepa Babin @TMC Kollam antibody + RBC w/ antigen
  6. 6. Incomplete Antibodies Deepa Babin @TMC Kollam + RBC w/ antigen antibodies
  7. 7. <ul><li>ABO BLOOD GROUP </li></ul><ul><li>Rh BLOOD GROUP SYSTEM </li></ul><ul><li>OTHERS </li></ul><ul><li>A. MNS </li></ul><ul><li>B. I/i </li></ul><ul><li>C. DUFFY </li></ul><ul><li>D. KELL </li></ul><ul><li>E. KIDD </li></ul><ul><li>F. P </li></ul><ul><li>G. LUTHERAN </li></ul><ul><li>H. LEWIS </li></ul>BLOOD GROUP SYSTEMS Deepa Babin @TMC Kollam
  8. 8. Deepa Babin @TMC Kollam
  9. 9. ABO BLOOD GROUP <ul><li>First blood groups discovered-Landsteiner(1900) </li></ul><ul><li>Most significant for transfusion practice </li></ul><ul><li>ABO compatibility is essential before other pretransfusion test is performed </li></ul><ul><li>ABO antigens are the only Ags for which reciprocal antibodies consistently and predictably exist in serum of normal individuals </li></ul>Deepa Babin @TMC Kollam
  10. 10. ABO BLOOD GROUP Deepa Babin @TMC Kollam Anti-A Anti-B Anti-A,B none O Blood Group RBC Ag A A B B AB A & B O none
  11. 11. <ul><li>GROUP A </li></ul><ul><ul><li>Express A antigen on RBC surface </li></ul></ul><ul><ul><li>Genotypes AA or AO </li></ul></ul><ul><ul><li>Have naturally occurring, clinically significant, predominantly IgM (with a small amount of IgG) antibodies against type B (anti-B) </li></ul></ul><ul><ul><li>Subgroups </li></ul></ul><ul><ul><ul><li>A1 (80%) </li></ul></ul></ul><ul><ul><ul><li>A2 (20%) </li></ul></ul></ul><ul><ul><ul><li>Significance: some with A2 have antibodies against the A1 subgroup (anti-A1) </li></ul></ul></ul>ABO BLOOD GROUP… Deepa Babin @TMC Kollam
  12. 12. ABO BLOOD GROUP… <ul><li>GROUP B </li></ul><ul><ul><li>Express B Ag on RBC surface </li></ul></ul><ul><ul><li>Genotypes BB or BO </li></ul></ul><ul><ul><li>Have naturally occurring clinically significant, predominantly IgM (with a small amount of IgG) antibodies against type A cells </li></ul></ul>Deepa Babin @TMC Kollam
  13. 13. ABO BLOOD GROUP… <ul><li>GROUP O </li></ul><ul><ul><li>Have neither A nor B antigens on their RBC </li></ul></ul><ul><ul><li>Genotype OO (“universal donors”) </li></ul></ul><ul><ul><li>Have naturally occurring, clinically significant, very high titer, anti-A, anti-B and anti-A,B antibodies </li></ul></ul><ul><ul><li>Maternal anti-A,B can cross the placenta to cause hemolytic disease of the newborn </li></ul></ul><ul><ul><li>Group O cells have the most H antigen </li></ul></ul>Deepa Babin @TMC Kollam
  14. 14. ABO BLOOD GROUP… <ul><li>GROUP AB </li></ul><ul><ul><li>Express A and B Ag on RBC surface </li></ul></ul><ul><ul><li>Genotypes A1B or A2B </li></ul></ul><ul><ul><li>have no ABO antibodies ( “universal recipients”) </li></ul></ul>Deepa Babin @TMC Kollam
  15. 15. H Antigen <ul><li>Seen on RBC of ABO-Universal distributed </li></ul><ul><li>BOMBAY (O h )PHENOTYPE </li></ul><ul><ul><li>Patients lack the H gene and therefore cannot make H antigen, A or B antigen on their red cells </li></ul></ul><ul><ul><li>Have very strong anti-A, anti-B, and anti-H and can only receive cells from a Bombay donor </li></ul></ul>Deepa Babin @TMC Kollam
  16. 16. ABO BLOOD GROUPS Deepa Babin @TMC Kollam Serum Ab RBC Ag Blood Group Anti-B A A Anti-A B B none A & B AB Anti-A Anti-B Anti-A,B none O
  17. 17. Blood being tested Typing Serum Type AB (contains agglutinogen A and B ) Type B (contains agglutinogen B) Type A (contains agglutinogen A) Type O (contains no agglutinogen) Anti – A Anti – B red cells Deepa Babin @TMC Kollam
  18. 18. ABO BLOOD GROUP… Testing <ul><li>REVERSE GROUPING /TYPING </li></ul><ul><ul><li>Also called “serum” or “back” typing </li></ul></ul><ul><ul><li>Uses patient’s serum versus commercial A1 and B cells </li></ul></ul><ul><ul><li>Analyzes patient’s serum for the presence of anti-A and anti-B antibodies </li></ul></ul>Deepa Babin @TMC Kollam
  19. 19. ABO BLOOD GROUP… Testing <ul><li>Testing for antibodies is not reliable until 6 months of age </li></ul><ul><ul><li>Newborn serum may contain maternal IgG anti-a or anti-B </li></ul></ul>Deepa Babin @TMC Kollam
  20. 20. Deepa Babin @TMC Kollam
  21. 21. Rh BLOOD GROUP … <ul><li>Complex blood group with >50 described antigens </li></ul><ul><li>Rhesus monkey </li></ul><ul><li>D Ag is more potent </li></ul><ul><li>No natural AB against Rh </li></ul><ul><li>Nomenclature systems </li></ul><ul><ul><li>Fisher-Race (English) </li></ul></ul><ul><ul><li>Wiener (American) </li></ul></ul>Deepa Babin @TMC Kollam
  22. 22. Rh Groups: Fisher-Race Anti-e Anti-C Anti-D e C D 5 major antigens: D, C, E, c and e Deepa Babin @TMC Kollam genes linked closely- e C D Ab Ag Chromosome
  23. 23. Rh Groups: Weiner Rh 1 hr” Anti- hr’ hr’ Anti-rh” rh” Anti-rh’ rh’ Anti-Rho Rho Anti- hr” 5 major antigens: Rh o , rh’, rh”, hr’, hr” Deepa Babin @TMC Kollam Ab Ag CHROMOSOME R 1 single gene
  24. 24. Rh BLOOD GROUP… Testing <ul><li>Testing for D(Rh o ) is the most common Rh test performed </li></ul><ul><li>D antigens are potent immunogens. </li></ul><ul><ul><li>Of D-negative patients, 80% will develop an anti-D when transfused with a single unit of D-positive blood. </li></ul></ul>* Rh-positive simply means “D positive” Deepa Babin @TMC Kollam
  25. 25. Other blood group systems: <ul><li>Kell (K,k,Kx) </li></ul><ul><li>Duffy (Fy) </li></ul><ul><li>Kidd (Jk a , Jk b ) </li></ul><ul><li>MNSs </li></ul><ul><li>P </li></ul><ul><li>Lewis (Le) </li></ul><ul><li>Lutheran (Lu) </li></ul>Deepa Babin @TMC Kollam
  26. 26. Importance of other blood groups: <ul><li>May cause transfusion reactions or HDN </li></ul><ul><li>May interfere with crosshatching </li></ul><ul><li>For medico legal parenthood problems </li></ul>Deepa Babin @TMC Kollam
  27. 27. Deepa Babin @TMC Kollam
  28. 28. COMMON TESTS USED IN IMMUNOHEMATOLOGY <ul><li>Coombs Test (Antiglobulin Test) </li></ul><ul><li>RBC typing </li></ul><ul><li>Crossmatching </li></ul><ul><li>Antibody Screening </li></ul><ul><li>Compatibility Testing </li></ul>Deepa Babin @TMC Kollam
  29. 29. I . ANTIGLOBULIN (COOMB’S) TEST <ul><li>Remains the most important single test in Ab detection </li></ul><ul><li>Principle: </li></ul><ul><ul><li>Red blood cells sensitized by IgG or complement can be made to agglutinate by adding antihuman globulin </li></ul></ul>Deepa Babin @TMC Kollam
  30. 30. I. ANTIGLOBULIN (COOMB’S) TEST <ul><li>DIRECT COOMB’S TEST (DAT) </li></ul><ul><ul><li>Detects RBCs that have already been sensitized with IgG </li></ul></ul><ul><ul><li>Demonstrates that in vivo coating of RBC by Ab has occurred but does NOT identify the antibody </li></ul></ul>Deepa Babin @TMC Kollam
  31. 31. Deepa Babin @TMC Kollam
  32. 32. <ul><li>INDIRECT COOMB’S TEST (IAT) </li></ul><ul><ul><li>Detects antibodies to RBC antigens present in the patient’s serum </li></ul></ul><ul><ul><li>Detects in vitro red cell sensitization if red cells contain antigen corresponding to serum antibody </li></ul></ul><ul><ul><li>Procedure: </li></ul></ul><ul><ul><ul><li>STEP 1: </li></ul></ul></ul><ul><ul><ul><li>patient’s serum (with unknown Ab) + </li></ul></ul></ul><ul><ul><ul><li>RBC (with known Ag) </li></ul></ul></ul><ul><ul><ul><li>STEP 2: product of step 1 + Coomb’s reagent </li></ul></ul></ul>I . ANTIGLOBULIN (COOMB’S) TEST Deepa Babin @TMC Kollam
  33. 33. Deepa Babin @TMC Kollam
  34. 34. ANTIGLOBULIN TESTING Deepa Babin @TMC Kollam DIRECT ANTIGLOBULIN TEST (DAT) Patient’s red cells INDIRECT ANTIGLOBULIN TEST (IAT) Patient’s serum Detects in vivo antibody coating (sensitization) of red cells Detects in vitro red cell sensitization if red cells contain antigen corresponding to serum antibody <ul><li>Useful in : </li></ul><ul><li>Detection of hemolytic disease of the newborn (employing infant’s red cells) </li></ul><ul><li>Investigation of transfusion reactions </li></ul><ul><li>Detection of autoimmune hemolytic anemia (AIHA) </li></ul><ul><li>Detection of red cell sensitization by drugs (penicillin, cephalothin, alpha-methyldopa) </li></ul><ul><li>Useful in: </li></ul><ul><li>Detection and identification of unexpected antibodies </li></ul><ul><li>Compatibility testing (cross-matching) </li></ul><ul><li>Red cell antigen phenotyping </li></ul><ul><li>Investigation of transfusion reactions </li></ul><ul><li>Detecting Du antigen (weak D) </li></ul>
  35. 35. II.RBC typing <ul><li>Forward typing </li></ul><ul><li>Reverse typing </li></ul><ul><li>D antigen </li></ul>Deepa Babin @TMC Kollam
  36. 36. III.CROSSMATCHING <ul><li>Absence of agglutination or hemolysis is essential to the safety of blood transfusions </li></ul><ul><li>Agglutination or hemolysis in any phase of the transfusion (ie incompatibility) = presence of Ab and its corresponding Ag </li></ul>Deepa Babin @TMC Kollam
  37. 37. III.CROSSMATCHING <ul><li>Uses </li></ul><ul><li>To detect antibodies in the donor or recipient </li></ul><ul><li>To detect ABO typing discrepancies </li></ul>Deepa Babin @TMC Kollam
  38. 38. IV. ANTIBODY SCREEN <ul><li>Use: </li></ul><ul><ul><li>to demonstrate unexpected antibodies in the serum of the recipient that may destroy donor RBCs that were thought to be compatible on the basis of the Rh and ABO typing </li></ul></ul><ul><ul><li>Has replaced minor cross matching </li></ul></ul>Deepa Babin @TMC Kollam
  39. 39. V. PRE-TRANSFUSION / COMPATIBILITY TESTING <ul><li>Review of recipient’s blood bank history </li></ul><ul><li>ABO and Rh typing of recipient & donor </li></ul><ul><li>Antibody screening of recipient & donor serum </li></ul><ul><li>Major crossmatching </li></ul>Deepa Babin @TMC Kollam
  40. 40. Deepa Babin @TMC Kollam
  41. 41. HEMOLYTIC DISEASE OF THE NEWBORN (HDN) <ul><li>Also referred to as erythroblastosis fetalis </li></ul><ul><li>Occurs when the mother is alloimmunized to antigen(s) found on the RBC of the fetus </li></ul><ul><li>Destruction of fetal RBCs by mother’s IgG antibodies </li></ul>Deepa Babin @TMC Kollam
  42. 42. HEMOLYTIC DISEASE OF THE NEWBORN (HDN) <ul><li>Rh Incompatibility Rh (-) mother & Rh (+) baby </li></ul><ul><li>ABO Incompatibility “O” mother & “A” or “B” baby </li></ul>Deepa Babin @TMC Kollam
  43. 43. HDN DUE TO RH INCOMPATIBILITY <ul><li>Set-up: Rh(-) mother + Rh(+) baby </li></ul><ul><li>Rh (-) person exposed to Rh(+) blood will develop reaction after 2 – 4 weeks </li></ul><ul><li>Mother develops antibody against the Rh(+) blood coming from the baby </li></ul><ul><li>First baby is not affected; HDN occurs during subsequent pregnancies </li></ul>Deepa Babin @TMC Kollam
  44. 44. Deepa Babin @TMC Kollam Rh(-) RBC Rh(+) RBC
  45. 45. Deepa Babin @TMC Kollam Antibody
  46. 46. Deepa Babin @TMC Kollam
  47. 47. anemia hemoglobin degradation extramedullary hematopoiesis hydrops hypoxic injury to liver & heart  bilirubin jaundice kernicterus hepato- splenomegaly HEMOLYSIS Deepa Babin @TMC Kollam
  48. 48. Deepa Babin @TMC Kollam
  49. 49. Kernicterus Hydrops Deepa Babin @TMC Kollam
  50. 50. HDN DUE TO RH INCOMPATIBILITY <ul><li>May be prevented by giving anti-Rh to Rh(-) mother in the ante-natal (28 weeks) & immediate postnatal period (within 72 hours after delivery) </li></ul>Deepa Babin @TMC Kollam
  51. 51. ABO Hemolytic disease <ul><li>Maternofetal ABO incompatibility </li></ul><ul><li>A &B got IgM in blood which cannot cross placenta </li></ul><ul><li>O gp IgG-CROSS PLACENTA-HEMOLYTIC DISEASE </li></ul>Deepa Babin @TMC Kollam
  52. 52. COMPLICATIONS OF BLOOD TRANSFUSION <ul><li>Immunological/Non immunological </li></ul><ul><li>Infectious parameters(HIV,HBV,HCV,VDRL) </li></ul><ul><li>CMV,LEPTO ,MALARIA,TOXO,LEISH) </li></ul><ul><li>AUTOLOGUS BLOOD TRANSFUSION </li></ul>Deepa Babin @TMC Kollam
  53. 53. <ul><li>Programme created for Basic Vision on Immunohematology </li></ul><ul><li>Deepa Babin * </li></ul>Deepa Babin @TMC Kollam