Get Adobe Flash player


Аuthors: G. Myronenko, R. Pavliuk

Pages: 36–45



Introduction. The presence of antierythrocyte antibodies is one of the key elements in the diagnosis of autoimmune hemolytic anemia (AIHA). 80 % of AIHA are secondary (symptomatic) processes and they are a manifestation of underlying diseases (neoplasms, іnfections, lymphoproliferative disorders, autoimmune disorders, viral infections, immune deficiency states, medications and toxic effects). At the same time the traditional tests for the detection of antibodies (conglutination reaction with the adding of gelatin and Coombs test with polyspecific antiglobulin reagent) do not reveal the full range of antibodies which does not allow to form а sufficient clinician idea of the autoimmune nature of the hemolytic process and determine its activity for the prediction and optimization of patient’s treatment. This is a basic element of AIHA diagnostics detection of antierythrocyte antibodies. Due to current achievements in immunohaematology it is a necessary additional examination of patients.

Purpose. The purpose of current research study was to demonstrate the diagnostic and prognostic value range of modern immunohematological tests to evaluate autoimmune hemolytic processes.

Materials and methods. During current research 67 patients with acquired hemolytic anemia with positive tests serum for erythrocyte autoantibodies and varying degrees of severity of clinical and laboratory signs of hemolytic process were examined. The hemolysis in 1/3 patients had the secondary (symptomatic) origin – as a complication of malignant diseases of the blood, pernicious anemia, systemic connective tissue diseases, infections, uncontrolled unauthorized medication, toxic effects. We used the gel test for identification of erythrocytic antibodies, classes, subclasses and determining their density on erythrocytes as well as bringing the complement system (BioRad, USA). The statistical analysis using the statistical software package StatSoft STATISTICA 10.0.1011(USA) was performed in the research.

Results and Discussion. The results of the immunohaematological study shows a serologic range of autoantibodies: warm-active agglutinins – 52,2 %, cold-active agglutinins – 25,4 %, mixed cold- and warm-active antibodies – 7,5 %, hemolysins – 14,9 %. High-density warm agglutinins IgG1/IgG3 on the surface of red blood cells were accompanied by the maximum activity of the immune destruction and caused a significant impact on the progression of anemia – in terms of hemoglobin (p < 0,05) and the number of red blood cells (p < 0,01). Wide thermal amplitude of cold agglutinins (4–32 °C) was more important for the progression of hemolysis than their high titer. The combination of warm agglutinins IgG, IgA, IgM appeared to combine with a poor prognosis for the course of hemolytic process.

Conclusions. Coombs test is only a primary link in the diagnosis of AIHA. Consequently, further serological blood testing for the detection of all variants antibodies for the purpose of future diagnosis is needed. It is necessary to determine the density of antibodies on the surface of red blood cells in the case of warm IgG-antibodies and intensity of Coombs test reaction (3+/4+) and to examine the thermal amplitude and titer in case of identification of cold agglutinins.

Keywords: аutoimmune hemolysis, agglutinins, hemolysins, the density of antibodies.

This email address is being protected from spambots. You need JavaScript enabled to view it.

The full text

To view the full text


  1. Greer JP, Arber DA., Glader B, List AF, Means RT, Paraskevas F, Rodgers GM [editors]. Wintrobe's clinical hematology. 13th edition. Philadelfia: LIPPINCOTT WILLIAMS & WILKINS Publ., 2014. pp. 746-808.
  2. Guseva SA, Goncharov YaP. [Autoimmune hemolytic anemia: diagnosis and treatment (lecture)]. Ukrainian Journal of Hematology and Transfusiology. 2012; 1: 34-50.
  3. Arndt PA, Leger RM, Garratty G, Arndt PA. Serologic characteristics of ceftriaxone antibodies in 25 patients with drug-induced immune hemolytic anemia. Transfusion. 2012; 52(3): 602-612.
  4. Leger RM, Garratty G. Antibodies to oxaliplatin, a chemotherapeutic, are found in plasma of healthy blood donors. Transfusion. 2011; 51(8): 1740-1744.
  5. Arndt PA, Garratty G, Wolf CF, Rivera M. Haemolytic anaemia and renal failure associated with antibodies to trimethoprim and sulfamethoxazole. Transfus. Med. 2011; 21(3): 194-198.
  6. Lai M, Leone G, Landolfi R. Autoimmune hemolytic anemia with gel-based immunohematology tests. 2013; 139(4): 457-463.
  7. Lai M, De Stefano V, Landolfi R. Haemoglobin levels in autoimmune haemolytic anaemias at diagnosis: relationship with immunoproteins on red blood cells. 2014; 60(1): 127-131.
  8. Mineyeva NV. Grupy krovi cheloveka. Osnovy immunogematologii. 2nd [Human blood groups. Basics of immunohematology]. SPb.: A-print
  9. Pavliuk RP, Tymoshenko UV, Myronenko GA, Lavrovska LN, Sposib vyznachennya monophasnykh autoimunnykh kyslotnykh teplovykh ta kholodovykh hemolizyniv mikrometodom [Micromethod for determining the monophasic autoimmune acid heat and cold hemolysins]. Ukrainian patent, no.99298, 2015.
  10. Swiecicki PL, Hegerova LT, Gertz MA. Cold agglutinin disease. Blood.2013;122(7):1114-1121. Retrieved from:
  11. Berentsen S, Tjønnfjord GE.Diagnosis and treatment of cold agglutinin mediated autoimmune hemolytic anemia. Blood. Rev. 2012;26(3):107-115doi: 10.1016/j.blre.2012.01.002.
  12. Tymchenko AS, Yavorskyi VV, Maligon OI. Natsionalne kerivnytstvo z vyrobnychoi transfuziologii dlya zakladiv, pidrozdiliv ta laboratory sluzhby krovi [National guidelines on the production of transfusion for institutions, departments and laboratories blood service]. Kharkiv: Zoloti storіnki
  13. Chaudhary RK, Sudipta Sekhar Das. Autoimmune hemolytic anemia: From lab to bedside. Asian J. Transfus Sci. 2014; 8(1): 5–12. doi: .
  14. BarcelliniW,Fattizzo B, Zaninoni A, Radice T, NicheleI, Di Bona E, LunghiM, TassinariC, AlfinitoF, FerrariA, LeporaceAP, Niscola P,CarpenedoM, BoschettiC, Revelli N, Maria МА, Consonni D, Scaramucci L, De Fabritiis P, Tagariello G, Gaidano G, Rodeghiero F, Cortelezzi A, Zanella A. Clinical heterogeneity and predictors of outcome in primary autoimmune hemolytic anemia: a GIMEMA study of 308 patients. Blood.2014; 124(19): 2930-2936. doi: 10.1182/blood-2014-06-583021.
  15. Bodivit G, Bensussan A, Fournie J, Yamada K, Copie-Bergman C, Bodivit G, Bensussan A, ean- Fournie JJ, Godeau B, Bierling P, Izui S. IgA-mediated human autoimmune hemolytic anemia as a result of hemagglutination in the spleen, but independently of complement activation and Fc-RI. Blood. 2010; 116(20): 4141-4147.