Lymphoproliferative disease with IgM lambda monoclonal protein and autoimmune hemolytic anemia. A report of four cases and a review of the literature.

Four patients with lymphoproliferative disease with immunoglobulin M lambda (IgMlambda) monoclonal proteins and severe autoimmune hemolytic anemia are described. These patients had many features in common that may warrant their recognition as a specific entity within the lymphoproliferative spectrum. In each case, a wide thermal range low titer cold agglutinin was present. The association of cold autoimmune hemolytic anemia with IgMlambda monoclonal protein and lymphoproliferative disease is unusual. The literature on IgM monoclonal proteins associated with lymphoproliferative disease is reviewed with emphasis on the presence of direct antiglobulin test positive autoimmune hemolytic anemia.     

Complement Activation by Cold Agglutinins

Purified monoclonal human IgM cold agglutinins (CA) of different specificities (anti-I, anti-i, anti-Pr) were investigated for their complement-activating capacity in a homologous system. Incubation of human RBC with excess of IgM CA in the cold, and subsequently with human serum at 37 degrees C, resulted in striking differences in hemolysis. Hemolysis did not correlate to the amount of antibodies bound to RBC at 4 or 20 degrees C. Despite the hemolytic inefficiency of anti-i and anti-Pr CA tested, C1 fixation and subsequent activation of the classical pathway of complement could be assessed in all cases. Absolute numbers of C3 molecules bound to RBC, exceeding the critical level to initiate the terminal sequence of the complement cascade, could not fully explain the differences in the hemolytic activity of the CA. Since C8 binding protein (C8bp) carries I determinants it is hypothesized that anti-I-induced complement-mediated hemolysis might also be favored by the binding of the autoantibody to and probably steric hindrance of this major regulatory protein of the terminal complement sequence. The prominent role of homologous restriction of complement-mediated lysis as a protective mechanism can also be deduced from the fact that rabbit as well as rat serum as a source of heterologous complement lysed cold agglutinin-sensitized red blood cells more efficiently than human serum.     

Successful anti-CD20 monoclonal antibody treatment of severe autoimmune hemolytic anemia due to warm reactive IgM autoantibody in a child with common variable immunodeficiency

Autoimmune hemolytic anemia due to warm reactive IgM autoantibodies is unusual, severe, and often fails to respond to standard immunosuppressive therapies in both adults and children. A 6-year-old girl with common variable immunodeficiency had longstanding steroid dependent, splenectomy-unresponsive, warm IgM autoantibody-mediated autoimmune hemolytic anemia. Rituximab, a monoclonal antibody directed against CD20 antigen, was used to deplete B lymphocytes and reduce autoantibody production. She received a total of six doses of rituximab (375 mg/m2). Therapy was well tolerated, and B-lymphocytes were effectively depleted from the peripheral blood. The patient was completely tapered off glucocorticoids. The patient has remained off immunosuppressive agents for 16 months despite the return of B lymphocytes to the peripheral circulation. She continues to require IVIG. Early treatment with rituximab might be an option for patients with warm reactive IgM autoantibody-mediated autoimmune hemolytic anemia not responding to other treatments or experiencing untoward side effects from those treatments.     

Cold agglutinin autoimmune hemolytic anemia in nonhematologic malignancies.

Four patients with nonhematologic malignancies had the simultaneous finding of hemolytic anemia due to high-titer cold agglutinins. In each patient, the cold agglutinin had "anti-I" specificity and was of the IgM kappa immunoglobulin class. Although patients with hematologic malignancies not uncommonly have cold agglutinins, the association between these antibodies and nonhematologic malignancies is unusual.     

Autoimmune hemolytic anemia in association with monoclonal IgM(kappa) with anti-i activity.

A patient with a warm autoimmune hemolytic anemia with an immunoglobulin G (IgG) panagglutinin, also had monocional IgM(kappa) cold agglutinin with anti-i activity. Ninety per cent of the peripheral blood lymphocytes had surface immunoglobulin and the number of T cells was diminished. A subpopulation of the patient's lymphocytes formed rosettes with cord (i) erythrocytes and not with adult (l) erythrocytes. The finding of increased lymphocytes bearing i-binding sites and a monoclonal antibody with anti-i activity could be related to shared idiotypic determinants between antigen-binding sites and serum antibody. The occurrence of two autoantibodies in this patient suggests an immune regulatory disorder.     

Inefficacy of Plasma Exchange in Cold Agglutinin Hemolytic Anemia — A Case Study

A therapeutic trial of plasma exchange was conducted in an 82-year-old woman with severe cold agglutinin hemolytic anemia, who was unresponsive to conventional treatment. In spite of considerable improvement of pertinent parameters (total IgM, cold agglutinin titer and thermal amplitude) this treatment failed to induce a clinical remission. This failure may be the result of the ability of the cold agglutinin to fix complement to the red cell membrane at temperatures much higher than the thermal amplitude. A simple laboratory test to detect this dissociation between complement fixing ability and agglutination activity is described.     

Clinical and immunologic features of transient cold agglutinin hemolytic anemia

Atypical pneumonitis, often induced by infection with Mycoplasma pneumoniae, is frequently associated with elevated cold agglutinin titers but only rarely with significant hemolytic anemia. An example and documentation of the clinical and immunologic course of such transient cold agglutinin hemolytic anemia is presented, and the immunochemical characteristics of cold agglutinins in this syndrome are assessed in regard to their biologic implications and diagnostic significance. Transient cold agglutinins, such as observed in this case, commonly appear to be of a restricted polyclonal character and as first demonstrated in this case may possess structural determinants usually considered unique for monoclonal cold agglutinins. Although the immunopathogenetic mechanisms and certain clinical manifestations of the various forms of cold agglutinin hemolytic anemia appear similar, the immunogenic basis of cold agglutinin production and the molecular structure of transient cold agglutinins are quite distinctive and provide reasonably reliable guidelines for the differential diagnosis of the cold agglutinin syndromes and for consideration of appropriate clinical management.     

Beneficial Effect of Rituximab in Combination with Oral Cyclophosphamide in Primary Chronic Cold Agglutinin Disease

Cold agglutinin disease (CAD) is an uncommon autoimmune hemolytic anemia characterized by B-cell proliferation. Conventional therapies for primary CAD such as corticosteroids, oral alkylating agents, splenectomy, interferon alpha, and plasma exchange are often ineffective at controlling the disease. The anti-CD20 monoclonal antibody rituximab (MabThera) depletes B-lymphocytes and thereby interferes with the production of cold agglutinin. We describe an elderly patient with primary (idiopathic) chronic CAD refractory to steroids who was successfully treated with 4 weekly infusions (375 mg/m2) of rituximab and 6 months of oral cyclophosphamide at a dosage of 60 mg/m2 per day. The increase in hemoglobin level and the decline in the plasma cold agglutinin titer were rapid (from the second rituximab infusion). The hematologic remission persisted for at least 8 months after treatment start, with no adverse effects. Rituximab and cyclophosphamide may be supplementary therapeutic modalities whose combination warrants further clinical investigation.     

Complement inhibitors to treat IgM-mediated autoimmune hemolysis

Complement activation in autoimmune hemolytic anemia may exacerbate extravascular hemolysis and may occasionally result in intravascular hemolysis. IgM autoantibodies as characteristically found in cold autoantibody autoimmune hemolytic anemia, in cold agglutinin disease but also in a considerable percentage of patients with warm autoantibodies are very likely to activate complement in vivo. Therapy of IgM-mediated autoimmune hemolytic anemia mainly aims to decrease autoantibody production. However, most of these treatments require time to become effective and will not stop immediate ongoing complement-mediated hemolysis nor prevent hemolysis of transfused red blood cells. Therefore pharmacological inhibition of the complement system might be a suitable approach to halt or at least attenuate ongoing hemolysis and improve the recovery of red blood cell transfusion in autoimmune hemolytic anemia. In recent years, several complement inhibitors have become available in the clinic, some of them with proven efficacy in autoimmune hemolytic anemia. In the present review, we give a short introduction on the pathogenesis of autoimmune hemolytic anemia, followed by an overview on the complement system with a special focus on its regulation. Finally, we will discuss complement inhibitors with regard to their potential efficacy to halt or attenuate hemolysis in complement-mediated autoimmune hemolytic anemia.     

Treatment of autoimmune hemolytic anemias

Treatment of autoimmune hemolytic anemias varies depending on whether the patient has autoimmune hemolytic anemia of warm antibody type, cold agglutinin syndrome, paroxysmal cold hemoglobinuria, or autoimmune hemolytic anemia secondary to an underlying disorder. Initial therapy for warm antibody autoimmune hemolytic anemia should be corticosteroids, such as prednisone at conventional doses of 1 to 1.5 mg/kg/d orally. Criteria must be established to determine whether the therapeutic response is adequate, because long-term therapy may lead to significant detrimental side effects. Splenectomy has the advantage over therapeutic options in that it has the potential for complete and long-term remission. The major adverse effect is the syndrome of overwhelming postsplenectomy infection. Other therapeutic options, which are less likely to have long-term benefit, are immunosuppressive drugs, danazol, intravenous immunoglobulin, and plasma exchange. Therapy of cold agglutinin syndrome often is unsatisfactory. All patients should avoid exposure to cold, and if additional therapy is necessary, the therapies used for warm antibody autoimmune hemolytic anemia may be tried with less likelihood of response. Paroxysmal cold hemoglobinuria requires aggressive supportive therapy, generally supplemented by corticosteroids. Hemolysis usually terminates spontaneously. Patients with secondary autoimmune hemolytic anemia may be treated similarly to those with idiopathic autoimmune hemolytic anemia, and additional therapy for the underlying disorder also may result in remission of the hemolysis.     

Acute autoimmune hemolytic anemia due to a low molecular weight IgM cold hemolysin associated with episodic lymphoid granulomatous vasculitis

The case of a patient with a 3 year long illness characterized by episodic localized lymphadenitis and concurrent acute autoimmune hemolytic anemia is reported. Histopathologically the lymphoid disease was granulomatous necrotizing vasculitis of the Wegener type. The acute hemolysis occurring with these episodes was associated with extreme erythrophagocytosis in the peripheral blood without hemoglobinuria. The unique autoantibody resembled the cold agglutinins of lymphoproliferative diseases in that it was an immunoglobulin M (IgM) with only kappa light chains and had anti-I specificity. However, the antibody had no agglutinating ability, demonstrated biphasic thermal requirements for in vitro hemolysis (cold, then warm, incubation) and was of approximately 7S size. Despite the requirement for incubation at less than 10 °C to produce in vitro hemolysis, clinical episodes of severe hemolytic anemia were unassociated with cold exposure. An excellent response to cortico-steroids contrasted with the typical experience with cold-reacting red cell autoantibodies.     

成人支原体肺炎并发溶血性贫血一例并文献复习

目的 提高临床医生对成人支原体肺炎并发溶血性贫血的认识.方法 报道1例成人支原体肺炎并发溶血性贫血病例,以mycoplasma pneumonia、hemolytic anemia、adult及case report为检索词,检索时限为1967年1月至2011年3月,通过PubMed检索系统进行检索;同时以“肺炎,支原体,溶血性贫血,成人”为检索词,检索时限为1978年1月至2011年3月,通过万方数据库对中文文献进行检索,共检索出相关临床资料完整的病例报告9篇.结合国内外文献报道的9例同类病例的临床资料进行分析.结果 患者女,29岁,因“发热伴咳嗽、咳痰15 d,血红蛋白下降2d”入院.X线胸片示左下肺渗出性病变,发病第14天外周血血红蛋白下降,最低至83 g/L,网织红细胞为0.04,同时室温下抽出的静脉血在试管中凝集,血清支原体抗体IgM阳性,冷凝集素滴度为1∶256.经抗感染治疗2周后血红蛋白含量恢复正常,肺部病变消失,诊断为支原体肺炎并发溶血性贫血.复习国内外文献报道的9例临床资料完整的成年病例,10例中男6例,女4例,年龄29 ～60岁,8例在发病第8～18天出现溶血性贫血,10例均出现血冷凝集素滴度增高(其中7例＞1∶1024),2例发生血管内栓塞.10例中9例痊愈,1例死亡.结论 支原体肺炎并发溶血性贫血多发生在发病第8～18天,与冷凝集素滴度增高相关,部分病例同时出现血管内栓塞.多数病例预后良好,少数重症病例预后不良.     

A comparison of methods of monoclonal immunoglobulin quantitation

Quantitation of monoclonal immunoglobulins was carried out using the following methods: densitometric scanning of electrophoretograms on cellulose acetate gel, single radial immunodiffusion, nephelometry and, in monoclonal IgM, also sedimentation analysis. To compare the methods, 20 sera with monoclonal IgG, 16 sera with monoclonal IgA and 19 sera with monoclonal IgM were used. The methods correlated well in monoclonal IgM and IgG while in IgA correlation was found only between densitometry and radial immunodiffusion. Closest correlation was observed between densitometry and sedimentation analysis in quantifying monoclonal IgM (r = 0.954). Despite good correlation, the individual values of monoclonal immunoglobulins established by different methods showed marked differences. In comparison with the immunochemical methods, the quantitation of monoclonal immunoglobulins using densitometric scanning of electrophoretograms is accessible, quick and relatively more precise method.     

Complement deposition in autoimmune hemolytic anemia is a footprint for difficult-to-detect IgM autoantibodies

In autoimmune hemolytic anemia autoantibodies against erythrocytes lead to increased clearance of the erythrocytes, which in turn results in a potentially fatal hemolytic anemia. Depending on whether IgG or IgM antibodies are involved, response to therapy is different. Proper identification of the isotype of the anti-erythrocyte autoantibodies is, therefore, crucial. However, detection of IgM autoantibodies can be challenging. We, therefore, set out to improve the detection of anti-erythrocyte IgM. Direct detection using a flow cytometry-based approach did not yield satisfactory improvements. Next, we analyzed whether the presence of complement C3 on a patient’s erythrocytes could be used for indirect detection of anti-erythrocyte IgM. To this end, we fractionated patients’ sera by size exclusion chromatography and tested which fractions yielded complement deposition on erythrocytes. Strikingly, we found that all patients with C3 on their erythrocytes according to standard diagnostic tests had an IgM anti-erythrocyte component that could activate complement, even if no such autoantibody had been detected with any other test. This also included all tested patients with only IgG and C3 on their erythrocytes, who would previously have been classified as having an IgG-only mediated autoimmune hemolytic anemia. Depleting patients’ sera of either IgG or IgM and testing the remaining complement activation confirmed this result. In conclusion, complement activation in autoimmune hemolytic anemia is mostly IgM-mediated and the presence of covalent C3 on patients’ erythrocytes can be taken as a footprint of the presence of anti-erythrocyte IgM. Based on this finding, we propose a diagnostic workflow that will aid in choosing the optimal treatment strategy.     

Two new glucose 6-phosphate dehydrogenase variants associated with congenital nonspherocytic hemolytic anemia found in Japan: Gd(-) tokushima and gd(-) tokyo

Two new variants of glucose 6-phosphate dehydrogenase (G6PD) deficiency associated with chronic nonspherocytic hemolytic anemia were discovered in Japan. Gd(-) Tokushima was found in a 17-year-old male whose erythrocytes contained 4.4% of normal enzyme activity. Partially purified enzyme revealed a main band of normal electrophoretic mobility with additional two minor bands of different mobility; normal Km G6P, and Km NADP five- to sixfold higher than normal; normal utilization of 2-deoxy-G6P, galactose-6P, and deamino-NADP; marked thermal instability; a normal pH curve; and normal Ki NADPH. The hemolytic anemia was moderate to severe. Gd(-) Tokyo was characterized from a 15-year-old male who had chronic nonspherocytic hemolytic anemia of mild degree. The erythrocytes contained 3% of normal enzyme activity, and partially purified enzyme revealed slow electrophoretic mobility (90% of normal for both a tris-hydrochloride buffer system and a tris-EDTA-borate buffer system, and 70% of normal for a phosphate buffer system); normal Km G6P and Km NADP; normal utilization of 2-deoxy-G6P, galactose-6P, and deamino-NADP; greatly increased thermal instability; a normal pH curve; and normal Ki NADPH. These two variants are clearly different from hitherto described G6PD variants, including the Japanese variants Gd(-) Heian and Gd(-) Kyoto. The mothers of both Gd(-) Tokushima and Gd(-) Tokyo were found to be heterozygote by an ascorbate-cyanide test.     

Autoimmune thrombocytopenia in Waldenstr?m's macroglobulinemia.

Autoimmune phenomena are well-recognised complications of Waldenstr?m's macroglobulinemia (WM) and IgM monoclonal gammopathy. Peripheral neuropathy and cold agglutinin hemolytic anemia are the most common reported and occur in 5-10% of patients. Autoimmune thrombocytopenia has been rarely reported in WM and its incidence is not known. In this study we report the case of a 67-year-old man who presented with autoimmune thrombocytopenia who was subsequently found to have WM. Laboratory investigation demonstrated that platelet-associated IgM (PAIgM) but not PAIgG was clearly elevated compared to normal controls. In addition the patient's serum reacted strongly with a panel of donor platelets analysed with an indirect platelet immunofluorescence assay utilising an anti-IgM secondary antibody. Glycoprotein specificity could not however be demonstrated by ELISA techniques for platelet glycoproteins IIbIIIa, IaIIa, IbIXa, and IV. We also reviewed the case records of 104 additional cases of WM diagnosed at our institution between 5/93 and 5/99. Three further cases with clinically significant autoimmune thrombocytopenia were identified. The overall incidence of autoimmune thrombocytopenia (4/105, 3.8%) in this cohort of patients was similar to the incidence of peripheral neuropathy (7/105, 6.7%) and cold agglutinins (3/105, 2.9%).     

Hereditary Aspects of Autoimmune Hemolytic Anemia; a Retrospective Analysis

Summary. A family with three siblings involved in autoimmune hemolytic anemia is presented. One sibling had an idiopathic variety of the Evans' syndrome type of autoimmune hemolytic anemia. The other two siblings had secondary forms of autoimmune hemolytic anemia, with plasma cell myeloma present in one and systemic lupus erythematosus in the other. A retrospective analysis of the family histories of forty-four patients with idiopathic autoimmune hemolytic anemia revealed that 20 % had family members involved in clinically detectable autoimmune disease.
The hypothesis is presented that autoimmune hemolytic anemia results from a fundamental aberration of the immune apparatus which prevents the establishment of a normal immune homeostatic mechanism. This abnormality appears to be transmitted on a familial basis, and is not primarily related to erythrocyte immune homeostasis. As such, the concept of 'idiopathic' autoimmune hemolytic anemia may be illusory and simply represent a partial diagnosis of a more generalized medical disorder.     

Seroepidemiology of infection with Toxoplasma gondii in healthy blood donors of Durango, Mexico

Background

Human parvovirus B19 is the etiologic agent of erythema infectiosum in children. It is also associated with other clinical manifestations in different target groups. Patients with chronic hemolytic anemia are at high risk of developing acute erythroblastopenia following infection by the virus. They usually become highly viremic and pose an increased risk of virus transmission. Close monitoring of such high risk groups is required for epidemiologic surveillance and disease prevention activities. Here we report a molecular epidemiological study on B19 virus infection in Tunisian patients with chronic hemolytic anemia.

Methods

This study was conducted on 92 young chronic hemolytic anemia patients who attended the same ward at the National Bone Marrow Transplantation Center of Tunis and 46 controls from a different hospital. Screening for IgM and IgG anti-B19 antibodies was performed using commercially available enzyme immunoassays and B19 DNA was detected by nested PCR in the overlapping VP1/VP2 region. DNA was sequenced using dideoxy-terminator cycle sequencing technology.

Results

Anti-parvovirus B19 IgG antibodies were detected in 26 of 46 sickle-cell anemia patients, 18 of 46 β-thalassemia and 7 of 46 controls. Anti-parvovirus B19 IgM antibodies were detected only in 4 of the sickle-cell anemia patients: two siblings and two unrelated who presented with acute erythroblastopenia at the time of blood collection for this study and had no history of past transfusion. B19 DNA was detected only in sera of these four patients and the corresponding 288 bp nested DNA amplicons were sequenced. The sequences obtained were all identical and phylogenetic analysis showed that they belonged to a new B19 virus strain of Genotype1.

Conclusion

A new parvovirus B19 strain of genotype1 was detected in four Tunisian patients with sickle-cell anemia. Virus transmission appeared to be nosocomial and resulted in acute erythroblastopenia in the four patients. The possibility of independent transmission of this B19 variant to the patients is unlikely in light of the present epidemiological data. However this possibility cannot be ruled out because of the low genetic variability of the virus.     

Evans syndrome associated with idiopathic mixed-type autoimmune hemolytic anemia

A 60-year-old man was admitted to our hospital with severe anemia and blood findings showed hemolytic anemia. Further serological examination revealed both warm-reactive autoantibody and cold agglutinin against erythrocytes. The cold agglutinin showed a low titer, 1 : 32 at 4 degrees C, and had a high thermal amplitude of 30 degrees C or higher, resulting in sufficient activity for hemolysis. Since no underlying disorders could be detected, the diagnosis was idiopathic mixed-type autoimmune hemolytic anemia. Although thrombocytopenia (Evans syndrome) subsequently appeared, corticosteroid was extremely effective for both anemia and thrombocytopenia. In this report we describe a rare case of Evans syndrome associated with mixed-type autoimmune hemolytic anemia, which had a dramatic response to corticosteroid therapy.     

Chronic hemolytic anemia due to a monoclonal IgG cold agglutinin with anti-Pr specificity

Immunologic studies performed in a case of autoimmune chronic hemolytic anemia with low titer cold agglutinins (1/16) demonstrated that the cold agglutinins corresponded to monoclonal IgG with anti-Pr specificity. This antibody had a large thermal amplitude, being active at 37 degrees C. These unusual characteristics may define a distinct subset of chronic cold agglutinin disease.