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A modified test for Brucella agglutinins.

0.1% Protamine sulphate in normal (0.85% w/v) saline has been used as a diluent in Brucella serology instead of saline as such. The use of protamine sulphate in this concentration has obviated the need for performing the modified Coombs test to detect non-agglutinating Brucella antibodies. This method has been used in the Microbiological Diagnostic Unit of the University of Melbourne in the routine titration of 721 sera and has proved to give reliable and reproducible results.

Agglutination Tests↗

Amiodarone-induced severe hepatitis mediated by immunological mechanisms.

A 64-year-old man developed a fulminant hepatitis 4 days after initiation of amiodarone treatment and a total dose of 7.1 g. The direct Coombs test was positive and became negative again soon after stopping treatment. Immediately after stopping treatment the extremely increased parameters of hepatic failure returned to normal again. A rechallenge with 200 mg of amiodarone was accompanied by a positive Coombs test which again became negative after several days. We conclude that the occurrence of an acute hepatitis soon after initiation of amiodarone treatment is mediated by immunological mechanisms. There should be high vigilance with respect to this rare life-threatening adverse drug reaction.

Amiodarone↗

Erythrocyte survival in patients with porcine xenograft aortic and mitral valves.

Ten patients with porcine xenograft aortic and mitral valve prostheses were studied three to 36 months (mean 15 months) postoperatively for evidence of hemolysis. Studies included complete blood count, reticulocyte count, red cell indices, percentage of schistocytes on blood smears, bilirubin concentration, lactic dehydrogenase, serum iron, total iron binding capacity, haptoglobin, serum folate and vitamin B12 levels, Coombs' test, methemoglobin reduction test, autologous 51Cr erythrocyte survival, and urinary examination for iron and hemosiderin. All patients were hemodynamically stable. Nine patients had normal valve function and no evidence of hemolysis. One patient with paravalvular aortic regurgitation had mechanical hemolytic anemia with a negative Coombs' test. Porcine valve xenografts do not seem to be associated with hemolysis unless complicated by a paravalvular leak.

Anemia, Hemolytic↗

Mycophenolate mofetil in the treatment of autoimmune HCV-associated haematological disorders showing steroid resistance or dependence.

We report two cases of hepatitis C virus (HCV) associated autoimmune haematological disorders successfully treated with an unusual protocol (mycophenolate mofetil: MMF). The first case was a male patient with chronic HCV infection who developed, during interferon (IFN)/ribavirin therapy, severe autoimmune thrombocytopenia unresponsive to steroids. MMF was then administered and, simultaneously, the steroid dose was gradually reduced until withdrawal. Following this strategy, a progressive increase in platelet count and complete negativity of anti-PLT antibodies were achieved without changes in HCV-RNA quantitative determination. The second case was a woman with HCV liver cirrhosis with severe anaemia and Coombs test positivity partially responsive to continuous administration of steroid high doses. However, this treatment unmasked a severely painful vertebral osteoporosis. For this reason we introduced MMF and simultaneously steroid therapy was progressively reduced until withdrawal. Haemoglobin reached a normal value and the Coombs test became negative within 60 days. These case reports suggest that MMF may represent an interesting therapeutic approach for autoimmune HCV associated haematological disorders.

Aged↗

Scleroderma associated with anemia and thrombocytopenia that responded well to cyclosporin.

A 55-year-old woman came to our hospital because of cutaneous sclerosis of the limbs in September 1996, and was diagnosed with scleroderma based on a skin biopsy. In August 1997, the cutaneous sclerosis became progressive (hemoglobin level, 4.3 g/dl; platelet count, 7 x 10(9)/l). The laboratory results were positive for the direct Coombs test, bone marrow aspiration showed a dry tap, and the bone marrow biopsy showed marked fatty marrow. Indium-111 bone marrow scintigraphy showed a markedly decreasing uptake. These findings indicated bone marrow hypoplasia associated with hemolytic anemia. After prednisolone therapy (60 mg) was initiated, the direct Coombs test became negative but the blood cell count did not increase. Then, 300 mg of cyclosporin was initiated and anemia and thrombocytopenia improved. The cyclosporin dosage was gradually decreased and the patient's hematological condition was good, although the cutaneous sclerosis changed only a little. This is a rare and interesting case of a patient with scleroderma associated with bone marrow insufficiency and hemolysis who responded well to cyclosporin.

Anemia, Hemolytic↗

Rebound hyperbilirubinemia in term infants after phototherapy.

OBJECTIVE: To determine whether a rebound in serum bilirubin level occurs within 24 hours after discontinuation of phototherapy in term infants with hyperbilirubinemia. METHODS: A retrospective medical record review of term infants with hyperbilirubinemia requiring phototherapy who were admitted over 24 months, June 1999 to December 2001, at King Abdul-Aziz University Hospital, Jeddah, Kingdom of Saudi Arabia was completed. Total serum bilirubin levels (TSB) at and up to 24 hours after termination of phototherapy (follow-up levels) were recorded. The difference between mean TSB level at termination of phototherapy and at follow-up was calculated using the paired t test. RESULTS: Three hundred and one infants, 53% (n=161) were boys, mean standard deviation (SD) for gestational age was 39.4 +/- 1.4 weeks, mean birth weight was 3200 +/- 600 g, mean TSB SD at termination of phototherapy was 193 +/- 46 micromole/liter (micromol/l) and at follow-up was 188 +/- 45 micromol/l for all infants with either positive or negative direct coombs' test results. The difference in mean TSB level at discontinuation of phototherapy and at follow-up was statistically significant, (t=3.12, P=0.002). The mean TSB level at termination of phototherapy was 179 +/- 47 micromol/l and at follow-up was 177 +/- 47 micromol/l for 65 infants with positive direct coombs' test results. This difference was statistically insignificant (t=0.725, P=0.47). The mean time interval between the termination of phototherapy and the measurement of follow-up TSB level was 8.3 +/- 5.3 hour. CONCLUSION: The rebound of bilirubin level after termination of phototherapy in otherwise healthy term infants is minimal; thus measurement of serum bilirubin is not required after termination of phototherapy and adds unnecessary expense, prolongs hospitalization, or both.

Female↗

[A delayed hemolytic transfusion reaction due to irregular antibodies with an anti-Kidd(a) specificity].

HISTORY AND CLINICAL FINDINGS: A 56-year-old woman was admitted for replacement of a previously implanted right hip prosthesis which had become loose (no infection). She limped painfully on a shortened leg whose mobility was markedly impaired. At the site of the previous operation the pelvis was lower by about 2 cm; the scar looked well healed. INVESTIGATIONS: Routine laboratory tests were normal except for moderately raised erythrocyte sedimentation rate (18/ 27 mm). There was complete loosening of the prosthesis on X-ray. TREATMENT AND COURSE: Intraoperative bleeding from a branch of the inferior gluteal artery required blood transfusion and further erythrocyte infusions became necessary. Jaundice developed on the 4th postoperative day and 7 days later the direct Coombs test was positive with demonstrable agglutination. Free irregular erythrocytic anti-Kidd(a) (anti-jk[a]) antibodies were found in the serum. To counteract the delayed haemolytic transfusion reaction, exclusively jk(a)-negative erythrocytes were infused. The jaundice gradually disappeared and the bilirubin values became normal. CONCLUSION: Jaundice and signs of haemolysis after erythrocyte transfusion may be due to delayed transfusion reaction and should be investigated with the direct Coombs test.

Antibodies↗

[Standard methods of blood group serology--state and trends of development].

The notion of transfusion serology comprises also serologic examinations in connection with transfusions of thrombocytes and leucocytes. The minimum demands of serological methods in the German Pharmacopoeia 7 (DAB 7) obligatory nowadays before transfusions of full blood and concentrates of erythrocytes should be changed and enlarged as follows: Obligatory carrying-through of the bed-side-test, inclusion of the ferment- and/or Coombs-test as major test into the cross typing, abolition of the minor test. In connection with the transfusions of leucocytes and thrombocytes determinations of the HLA-antibodies and cross typings are practicable in the institutions of the transfusion service. Transfusion must principally be striven for, taking into consideration the HLA-typing, but at present it is only limitedly possible. The search for donors among relations by blood increases the chances of the serological compatibility. Necessity and scale of an "emergency serology" still need discussion. Optimum haemotherapy is possible only then, when clinics and transfusion service work together in very close coordination.

ABO Blood-Group System↗

Hemolytic anemia associated with multiple autoantibodies and low serum complement.

A 37 year old woman with extravascular hemolytic anemia had a positive Monospot test associated with positive antiglobulin and anticomplement Coombs' tests, cold agglutinins and warm autoantibodies. IgG-kappa (k) antibodies, which reacted with all panel red cells at 37 degrees C, were eluted from her circulating red cells. However, neither immunoglobulins nor C3 was detected after her serum was adsorbed with heterologous red cell stroma at 37 degrees C and eluted at the same temperature in glycine buffer. In contrast, IgM-kappa and IgM-lambda (lambda), IgG-3-kappa, IgG4-lambda, IgA-lambda and C3 were eluted at 37 degrees C from heterologous red cell stroma after adsorption with her serum at 0 degrees C. Thus, antibodies of several types, which were present in the patient's serum, reacted optimally with red cell antigens at low temperature. Cold-reactive IgG3-kappa antibodies, which also capable of interacting with red cells at 37 degrees C, probably accounted for the IgG-kappa antibodies eluted from the patient's circulating red cells. The patient's serum C4 titers were decreased, with low normal to moderately depressed C3 and low normal C5, indicating that the anti-red cell IgM and/or IgG3-kappa antibodies probably fixed complement. A localized cold stress test resulted in a transient increase in plasma hemoglobin and a decrease in serum C3 titer. These findings, and the beneficial clinical response obtained with small doses of prednisone, suggest that both the cold-reactive antibodies and the IgG-kappa on circulating red cells were pathophysiologically significant. This is the first report of a patient with multiple red cell autoantibodies in whom serum complement component titers were determined in conjunction with characterization of the anti-red cell immunoglobulins. Subclinical infectious mononucleosis may have preceded the prolonged hemolytic episode. Clinical evidence of systemic lupus erythematosus has not appeared.

Adsorption↗

Immune abnormalities in myelodysplastic syndromes.

The immune states of 52 patients with myelodysplastic syndromes classified according to the FAB criteria were studied. Serum electrophoresis and immunoelectrophoresis, direct Coombs test, and tests for organ and non-organ specific antibodies were performed. Twenty six patients had immunoglobulin abnormalities: six (11.5%) had monoclonal gammopathy; 17 (32.6%) had polyclonal increases in serum immunoglobulin; while in three (5.8%) immunoglobulin concentrations were decreased. The distribution of immunoglobulin abnormalities among the five myelodysplastic syndrome subtypes was fairly uniform. Results of direct Coombs test were negative in all cases. Organ specific antibodies were not detected in any of the patients tested, although two patients were found positive for antinuclear antibodies. The presence of immunoglobulin abnormalities indicates an involvement of the lymphoplasmatic system in myelodysplastic syndromes.

Aged↗

Equine immune-mediated hemolytic anemia associated with Clostridium perfringens infection.

BACKGROUND: Previous studies have suggested an association between equine immune-mediated hemolytic anemia and clostridial infections or neck abscesses. OBJECTIVE: The purpose of this report was to describe and characterize the hematologic abnormalities in a horse with Clostridium-associated immune-mediated hemolytic anemia. We also retrospectively evaluated hematologic abnormalities in 8 horses with clostridial myositis or subcutaneous emphysema. METHODS: A 7-year-old Foreign Warm-Blood gelding was evaluated for anemia and a cervical abscess. CBCs and reticulocyte counts were obtained using an Advia 120 analyzer and evaluation of Wright's-stained smears. All cases of equine Clostridium spp.myositis or subcutaneous emphysema over a 7-year period were identified in a retrospective search of the University of Minnesota Veterinary Teaching Hospital database. Clinical, hematologic, and microbiological findings were recorded. RESULTS: Clostridium perfringens genotype A was isolated from the neck wound of the gelding. The CBC was characterized by severe regenerative anemia, intravascular hemolysis, and RBC agglutination. A direct Coombs' test was positive. Moderate numbers of spheroechinocytes were observed. The total automated reticulocyte count was 4.5% (56,700/microliter), with most reticulocytes having low absorbance (mature reticulocytes). The anemia responded to penicillin and steroidal and nonsteroidal immunosuppressive drugs. Of 8 horses with myositis, all of which involved the cervical region, 5 were anemic, 1 had a positive direct Coombs' test, and 2 had increased numbers of type III echinocytes and spheroechinocytes. CONCLUSIONS: The presence of type III echinocytes or spheroechinocytes may be helpful in diagnosing immune-mediated hemolytic anemia associated with clostridial infections in horses. Automated reticulocyte counts may detect very low levels of reticulocytosis in hemolytic anemia in horses.

Abscess↗

[A case of drug-induced immune hemolytic anemia during alpha-interferon therapy for renal cell carcinoma].

A Case of drug-induced immune hemolytic anemia during alpha-interferon (alpha-IFN) therapy for renal cell carcinoma is reported. A 61-year-old woman was admitted to Tochigi cancer center for the treatment of left renal cell carcinoma. She underwent left radical nephrectomy. From 7th post operative day, alpha-IFN (6 x 10(6) IU) was administered every other day. Diclofenac and indomethacin were administered for pain and high fever induced by alpha-IFN. Hemoglobinuria was first noted on 20th post operative day. Immune hemolytic anemia was suspected by blood examinations including Coombs' test, serum haptoglobin, serum LDH and serum GOT. alpha-IFN therapy and administration of diclofenac and indomethacin were discontinued and prednisolone therapy (50 mg daily) was begun. Hemoglobinuria disappeared by 5 days and laboratory data became in normal range by 3 weeks. Drug-induced immune hemolytic anemia was diagnosed by the process of laboratory data, especially of direct Coombs' test. Though drug-induced immune hemolytic anemia is a rare adverse side effect during alpha-INF therapy for renal cell carcinoma, it seemed an important complication.

Anemia, Hemolytic, Autoimmune↗

Neonatal hyperbilirubinemia and physical and cognitive performance at 17 years of age.

To estimate the effect of neonatal hyperbilirubinemia on long-term cognitive ability in full-term newborns with a negative Coombs test, we performed a 17-year historical prospective study of 1948 subjects. Intelligence tests and medical examinations performed at the military draft board were stratified according to serum bilirubin concentration. A logistic regression analysis was used to adjust for the confounding effects of gestational age, birth weight, Apgar score, ethnic origin, socioeconomic class, paternal education, birth order, and the administration of phototherapy and exchange transfusion. No direct linear association was shown between neonatal bilirubin levels and intelligence test scores or school achievement at 17 years of age. However, the risk for low intelligence test scores (IQ score less than 85) was found to be significantly higher (P = .014) among full-term male subjects with serum bilirubin levels above 342 mumol/L (20 mg/dL) (odds ratio, 2.96; 95% confidence interval, 1.29-6.79). This association was not observed among female subjects. We conclude that severe neonatal hyperbilirubinemia, among full-term male newborns with a negative Coombs test, could be associated with lower IQ scores at 17 years of age.

Adolescent↗

Outcome of alloimmunized fetuses managed solely by cordocentesis but not requiring antenatal transfusion.

A published algorithm for the frequency of fetal blood sampling in the management of fetal hemolytic disease allows many pregnancies to continue 1-3 months after the last sample until delivery at term. Though the positive predictive value for antenatal anemia is known, the likelihood of either neonatal hyperbilirubinemia or an unexpected anemia (< 30%) is not. The perinatal records of 51 antigen-positive neonates who did not require treatment antenatally were abstracted. As fetuses, these neonates had been prospectively coded as either low risk (pattern 1), moderate risk (pattern 2) or high risk (patterns 3 and 4) for antenatal anemia (hematocrit < 30%) based on their hematocrit, reticulocyte count, and the strength of the direct Coombs' test performed on their first sample. Delivery occurred at 38 +/- 2 weeks. Neonatal complications of hemolytic disease were common. Sixty-four percent required phototherapy, 17% one or more double-volume exchange transfusions, and 13% one or more simple transfusions for late-developing anemia. In all, 29% of neonates received postnatal transfusion therapy. The only correlation between the antenatal hematologic/serologic studies and the need for postnatal transfusion therapy was the strength of the indirect Coombs' test performed on the first fetal blood sample. Two neonates unexpectedly had anemia (4% risk). In the first, the hematocrit at 35 weeks was 40% and the ultrasound 1 week later normal. In one, the algorithm had been erroneously applied. Stability of the hematocrit in fetuses at risk to develop antenatal anemia can be accurately predicted by fetal blood tests performed weeks prior to delivery.(ABSTRACT TRUNCATED AT 250 WORDS)

Anemia, Hemolytic↗

[Red cell hypoplasia following autoimmune hemolytic anemia associated with T-CLL: report of a case and review of the literature].

A 78-year-old woman, who had axillary lymphadenopathy but no hepatosplenomegaly, was admitted because of lymphocytosis. The leukocyte count was 18.1 x 10(9)/l with 72% abnormal cells. Neither anemia nor thrombocytopenia was present. Many abnormal cells and erythroblasts were seen in the bone marrow. These abnormal cells had irregular nuclei but no granules in the cytoplasm. The surface markers of these cells were positive for E-rosette, CD 2, CD 3, and Leu 7 but negative for CD 4, CD 8, CD 11 (OKM 1), CD 16 (Leu 11), and HLA-DR. The DNA analysis revealed the rearrangement of T-cell receptor beta-chain genes. Direct Coombs test was positive and red-cell life-span (51Cr) was T 1/2 = 19.5 days. The patient was diagnosed as having T-CLL with mild autoimmune hemolysis and was followed without treatment. Seven months later, the leukemia cells of peripheral blood increased to 62.6 X 10(9)/l and the frank autoimmune hemolytic anemia developed. After prednisolone, vincristine and cyclophosphamide were administered, leukemia cells of blood decreased. Anemia with reticulocytopenia, however, persisted and direct Coombs test became negative. In the bone marrow at that time, many neutrophils and megakaryocytes besides leukemia cells were preserved, but erythroblasts were hardly seen, namely a pattern of red cell hypoplasia was observed. The patient deteriorated rapidly and died 26 months after initial recognition of lymphocytosis. When complement was added, the patient's serum obtained during red cell hypoplasia but not during autoimmune hemolysis inhibited BFU-E and CFU-GM in in vitro colony assays. This case indicates that not only B-CLL but also T-CLL is accompanied by immune hematocytopenia.

Aged↗

Secondary serological response of patients with chronic hepatosplenic suppurative brucellosis.

Chronic hepatosplenic suppurative brucellosis (CHSB) is a local reactivation of a previous brucellosis, coursing with an immunoglobulin G (IgG) and IgA secondary immunological response. The observation of two cases of CHSB with an apparent IgM response gave rise to a detailed serological study of three of our patients. We studied the first sample from all three patients and successive samples from two of them. In cases 1 and 2, we found samples with positive IgM lateral flow and IgM enzyme-linked immunosorbent assay results concomitantly with rheumatoid factor (RF); after absorption with anti-RF serum, these results were rendered negative. In patients 2 and 3 the diagnosis of brucellosis was delayed, because none of the test results were initially very significant. However, a clear seroconversion of IgG antibodies was observed in subsequent months; titers of the Brucellacapt and Coombs tests increased in similar ways, although Brucellacapt decreased more rapidly than Coombs, which persisted at high titers for years. In patient 3 a relapse was observed in the fourth year of follow-up, detected by Coombs and also by IgG lateral flow and counterimmunoelectrophoresis (CIEP), although not by the rose bengal, agglutination, or Brucellacapt tests. Serological changes in CHSB may sometimes be mild and are detected mainly by the Coombs test. Brucellacapt does not offer additional information, although IgG lateral flow and CIEP may be of some use. Careful surveillance of titer changes in the Coombs test is the best marker of infection activity. As the disease progresses, an intense IgG response may develop and RF sometimes appears, simulating an IgM response.

Adult↗

[Detection in bovine serum of non-agglutinating antibodies directed against occult antigens of Brucella abortus strain 19].

Injection of different animal species with particulate T-dependent antigens induces the appearance of two antibody populations belonging to IgG class, one of them with agglutinating activity and the other lacking this capacity, that is, non-agglutinating. In this paper, 3 adult female Hereford were inoculated subcutaneously, fortnightly, four times, with 5 x 10(10) heat killed smooth S19 Brucella abortus cells. Blood samples were taken ten days after the last inoculation and were individually processed. Serum agglutinating titres against the homologous antigen and the Coombs test score were similar, indicating that nonagglutinating antibody titres directed against surface antigens of the smooth phase were not significant (Table 1). When sera were incubated with a suspension of Brucella abortus S45/20 (a rough strain) they showed poor agglutinating ability, but they contained non-agglutinating activity at high level, as determined by the anti-gamma-globulin test (Table 1). Similar results were obtained when the same sera were adsorbed with Brucella abortus S19 up to elimination of the antibodies directed against surface antigens, and then incubated with a suspension of strain 45/20 (Table 1). These antibodies were isolated from sera by immunoadsorption, using a thick suspension of S45/20 and then dissociated with glycine-CHl buffer 0.1M pH 3. These antibodies showed little agglutinating capacity for S45/20; however non-agglutinating antibodies were bound to the bacteria up to 1.5 micrograms g as demonstrated by Coombs test. Apart from the incomplete antibodies against external antigens of smooth Brucella abortus strains detected in long-term stimulated cattle, non-agglutinating antibodies with specificity for deeper antigens of the smooth bacteria were evidenced.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Castleman's disease].

A 17-year-old girl developed acute colicky periumbilical pain with haematuria. On examination a tightly elastic space-occupying lesion about 10 cm in diameter was palpable around the umbilicus. Erythrocyte sedimentation rate (ESR) was raised to 113 mm in the first hour. There was a microcytic anaemia (8.7 g/dl) and a positive Coombs test. Ultrasonography revealed a periumbilical space-occupying lesion of decreased echogenicity and left hydronephrosis. Computed tomography demonstrated a homogeneous noninfiltrative tumour, about 7 cm in diameter, with increased contrast medium concentration. The tumour, which was covered by peritoneum and well circumscribed, was excised from the base of the mesentery. Histologically it proved to have hyaline-sclerotic changes in the blood vessels and lymphoid and plasma cellular infiltrations, corresponding to a mixed form of Castleman's disease. In a second operation a left nephroureterectomy was performed. The kidney, the regional lymph nodes and the tissue which had caused the stenosis of the ureter all showed the same changes as the tumour. Postoperatively the ESR and the blood count became normal, but the Coombs test remained positive. During a subsequent pregnancy the ESR again rose. The patient has remained symptom-free for 38 months after the second operation.

Adolescent↗