BACKGROUND: Detection methods for ABO antibody (Ab) titers vary across laboratories, and the results are different depending on the method used. We aimed to compare titer values using different detection methods for the measurement of ABO Ab titers. METHODS: For ABO Ab detection, pooled group A or B red blood cells (RBCs) were reacted with each of 20 sera from blood groups A, B, or O without dithiothreitol treatment. The room-temperature (RT) incubation technique and the indirect antiglobulin test (IAT) were used in the tube test and gel card test. Flow cytometry (FCM) was performed by using anti-IgM and anti-IgG Abs. RESULTS: Regardless of the blood groups tested, the FCM assay with anti-IgM showed the highest titer compared to the tube test and gel card test with RT incubation in both. The tube test with IAT showed a higher titer than the gel card test with IAT (Gel-IAT) or FCM with anti-IgG in blood group A and B, while Gel-IAT showed the highest titer relative to the other tests, only for the anti-A Ab in blood group O. CONCLUSIONS: There were significant differences in the titers depending on the detection method used, and each method showed a different detection capacity for each ABO Ab depending on the ABO blood group tested. Therefore, caution should be exercised in interpreting ABO Ab titer results, taking into consideration the detection method used and the blood group.
ABO Blood-Group System/*immunology ; Adult ; Aged ; Agglutination Tests/instrumentation/*standards ; Antibodies/*analysis ; Antibodies, Anti-Idiotypic/analysis ; Erythrocytes/chemistry/metabolism ; Female ; *Flow Cytometry ; Humans ; Male ; Middle Aged ; Temperature

OBJECTIVETo observe the clinical efficacy of Compound Xiatianwu tablets in the treatment of active rheumatoid arthritis.

METHODOne hundred and eighty cases with active rheumatoid arthritis were randomly divided into the control group (60 cases) with leflunomide, sulfasalazine, and celecoxib; the treatment group (120 cases) given compound Xiatianwu tablets on the basis of the control group, 2 tablets each time, 3 times/day, with the course of treatment of 3 month. Patients of the two groups were observed for clinical symptoms, erythrocyte sedimentation rate, C-reactive protein, rheumatoid factor, and immunoglobulin changes before and after the treatment.

RESULTThe treatment group showed an overall efficiency of 94. 2% , the Xiatianwu group showed an overall efficiency of 80. 0%, while the control group showed an overall efficiency of 81.7%. The difference among the three groups was statistically significant (P <0. 05 or P <0. 01) , indicating that the treatment group was superior to the Xiatianwu group, while the Xiatianwu group was superior to the control group.

CONCLUSIONCompound Xiatianwu tablets has remarkable effect in the treatment of active rheumatoid arthritis.

Adult ; Arthritis, Rheumatoid ; drug therapy ; immunology ; metabolism ; pathology ; C-Reactive Protein ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Erythrocytes ; drug effects ; Female ; Humans ; Immunoglobulins ; metabolism ; Male ; Middle Aged ; Tablets ; Treatment Outcome ; Young Adult

BACKGROUND: Validation of hemagglutination inhibition (HI) assays is important for evaluating antibody responses to influenza virus, and selection of erythrocytes for use in these assays is important. This study aimed to determine the correlation between receptor binding specificity and effectiveness of the HI assay for detecting antibody response to pandemic influenza H1N1 (pH1N1) virus. METHODS: Hemagglutination (HA) tests were performed using erythrocytes from 6 species. Subsequently, 8 hemagglutinating units of pH1N1 from each species were titrated by real-time reverse transcription-PCR. To investigate the effect of erythrocyte binding preference on HI antibody titers, comparisons of HI with microneutralization (MN) assays were performed. RESULTS: Goose erythrocytes showed most specific binding with pH1N1, while HA titers using human erythrocytes were comparable to those using turkey erythrocytes. The erythrocyte binding efficiency was shown to have an impact on antibody detection. Comparing MN titers, HI titers using turkey erythrocytes yielded the most accurate results, while those using goose erythrocytes produced the highest geometric mean titer. Human blood group O erythrocytes lacking a specific antibody yielded results most comparable to those obtained using turkey erythrocytes. Further, pre-existing antibody to pH1N1 and different erythrocyte species can distort HI assay results. CONCLUSIONS: HI assay, using turkey and human erythrocytes, yielded the most comparable and applicable results for pH1N1 than those by MN assay, and using goose erythrocytes may lead to overestimated titers. Selection of appropriate erythrocyte species for HI assay allows construction of a more reliable database, which is essential for further investigations and control of virus epidemics.
Adult ; Animals ; Antibodies, Neutralizing/immunology ; Antibodies, Viral/*analysis/immunology ; Chickens ; Erythrocytes/*metabolism ; Female ; Geese ; *Hemagglutination Inhibition Tests ; Horses ; Humans ; Influenza A Virus, H1N1 Subtype/genetics/immunology/*metabolism ; Influenza, Human/epidemiology/immunology/virology ; Male ; Middle Aged ; Neutralization Tests ; Pandemics ; Swine ; Turkeys

This study was purposed to investigate the efficacy of different whole flow lysis reagents for lysis of red blood cells in flow cytometric analysis. The expression of immunocytes was detected by flow cytometry after lysis of red blood cells using commercial reagents (Optilyse C, FACS Lysing Solution) and self-made red blood cell lysis reagents (RBC Lysis Buffer), the detection results were analyzed comparatively. The results showed that there was no significant difference in the percentage of CD3e(+), CD3e(+)CD4(+), CD3e(+)CD8a(+), CD3e(-)CD19(+), CD3e(-)NK1.1(+) and Gr-1(+) cells between 3 different lysis reagent groups. However OptiLyse C solution was suitable to Gr-1(+) cell detection, but did not suit to Foxp3(+) Treg detection. The self-made RBC Lysis Buffer and FACS Lysing Solution were suited to Foxp3(+) Treg detection. It is concluded that the use of self-made RBC Lysis Buffer for flow cytometry can get the lysis efficiency of commercially available lysis solutions when samples are prepared in accordance with standardized procedure. The self-made RBC Lysis Buffer not only can satisfy experimental requirements, but also can reduce the experimental costs.
Animals ; Erythrocyte Count ; Erythrocytes ; immunology ; metabolism ; Flow Cytometry ; instrumentation ; methods ; Immune System ; immunology ; Indicators and Reagents ; analysis ; Mice ; Mice, Inbred C57BL

OBJECTIVEThrough establishing different blood deficiency animal model, to evaluate enriching blood effect changes of the drug pair of Angelicae Sinensis Radix and Chuanxiong Rhizoma and each single herb, and to explore the effect characteristics of their compatibility.

METHODThree different methods of acetyl phenylhydrazine (APH) hemolytic method, cyclophosphamide (CTX) chemical damage method, APH-CTX complex method were used respectively to copy different blood deficiency model mice. Changes of orbit blood routine, thymus index, spleen index and ATPase activity of red cell membrane of model mice were tested.

RESULTCompared with normal group, all indexes had significant differences in three model mice. The drug pair and each single herb had significant impact on most indexes of the APH-CTX complex model mice, and on the individual indexes of APH hemolytic model mice and CTX chemical damage model mice. Therefore, APH and CTX complex blood deficiency model was more suitable for the enriching blood mechanism study of the drug pair of Angelicae Sinensis Radix and Chuanxiong Rhizoma. Compared with the single herb of Angelicae Sinensis Radix and Chuanxiong Rhizoma, the drug pair of them had presented enriching blood effect at different extent with strengthening trend in regulating the invigorating blood indexes, immune organs and energy metabolic enzymes.

CONCLUSIONThe results of this research have provided scientific basis for revealing the mutual promotive composition law of the drug pair of Angelicae Sinensis Radix and Chuanxiong Rhizoma, and responded effectively the mult-link and mult-target effect characteristics of Chinese medicine bio-effect, to offer reference for the bio-effect research of the complicated substance group of Chinese medicine and traditional Chinese medicine formulae, and to supply demonstrative reference for researching the formulae compatibility law which takes the single drug-drug pair-formulae as main line.

Administration, Oral ; Angelica sinensis ; chemistry ; Animals ; Ca(2+) Mg(2+)-ATPase ; drug effects ; metabolism ; Cyclophosphamide ; pharmacology ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; chemistry ; Erythrocytes ; drug effects ; enzymology ; Female ; Hematologic Diseases ; drug therapy ; etiology ; Hemoglobins ; drug effects ; Leukocytes ; drug effects ; Medicine, Chinese Traditional ; Mice ; Models, Animal ; Phenylhydrazines ; pharmacology ; Plant Roots ; chemistry ; Random Allocation ; Rhizome ; chemistry ; Sodium-Potassium-Exchanging ATPase ; drug effects ; metabolism ; Spleen ; drug effects ; immunology ; Thymus Gland ; drug effects ; immunology

OBJECTIVETo observe the effect of Shenfu Injection (SFI) on erythrocyte immunity function of patients undergoing cardiopulmonary bypass (CPB).

METHODSTwenty patients scheduled for valve replacement were randomly assigned to two groups, i.e. , the SFI group and the control group, 10 in each. SFI 1 mL/kg was intravenously dripped before induction of anesthesia and SFI 1 mL/kg administered in priming solution in the SFI group, while only normal saline was given to those in the control group. Venous blood samples (5 mL) were collected before induction of anesthesia (T1), 30 min CPB (T2), immediate by the end of CPB (T3), and postoperative 24 h (T4) respectively in all groups. The levels of the rosette rate of RBC-C3b receptor (RBC-C3bRR), the rosette rate of RBC-immune complex (RBC-ICR), plasma malondialdehyde (MDA), free hemoglobin (FHB), and interleukin-6 (IL-6) were detected.

RESULTSThere was no significant difference in the levels of RBC-C3bRR, RBC-ICR, plasma MDA, FHB, and IL-6 at T1 in both groups (P > 0.05). RBC-C3bRR at the rest time points was lower in the two groups than before induction of anesthesia. There was no statistical difference in FHB or IL-6 between T4 and T1 in the SFI group. The levels of RBC-ICR, MDA, FHB, and IL-6 increased in the two groups more than before induction of anesthesia at T2-4 ( P < 0.05). Besides, the RBC-C3b RR was lower, and levels of RBC-ICR, MDA, FHB, and IL-6 higher in the control group than in the SFI group, showing significant difference (P <0.05).

CONCLUSIONSFI could decrease the generation of inflammatory mediators during CPB, improve the erythrocyte immune function of patients during CPB, and reduce the risk of postoperative infection.

Adult ; Antigen-Antibody Complex ; blood ; Cardiopulmonary Bypass ; Drugs, Chinese Herbal ; pharmacology ; Erythrocytes ; drug effects ; immunology ; Female ; Hemoglobins ; analysis ; Humans ; Injections ; Interleukin-6 ; blood ; Male ; Malondialdehyde ; blood ; Middle Aged ; Receptors, Complement 3b ; metabolism

The aim of this study is to construct a bifunctional fusion protein, which can conjugate both human red blood cells and antibodies against classical swine fever virus (CSFV). We respectively amplified 2E8ScFv and mE2 genes from different recombinant vectors, in which 2E8ScFv gene is the single chain Fv gene against H antigen of human red blood cells, whereas mE2 gene is the main antigen coding region gene of CSFV E2 protein. We used overlap extension PCR to obtain an artificial fusion gene segment 2E8mE2 containing genes of Both 2E8ScFv and mE2, then ligated into the expression vector pET-DsbA and expressed in Escherichia coli BL21(DE3) PlysS host cells, after induced with IPTG the target fusion protein was successfully expressed and identified in inclusion bodies by SDS-PAGE and Western blotting. We purified the fusion protein and renatured it from inclusion bodies to obtain a native state of well biological activity. The Erythrocyte agglutination test results indicated that the fusion protein can conjugate both human red blood cells and antibodies of CSFV.
Erythrocytes ; immunology ; Escherichia coli ; genetics ; metabolism ; Humans ; Immunoglobulin Fragments ; biosynthesis ; genetics ; immunology ; Immunoglobulin Variable Region ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Single-Chain Antibodies ; biosynthesis ; genetics ; Viral Envelope Proteins ; biosynthesis ; genetics

In patients with hemolytic anemia associated with spherocytosis, differential diagnosis has to be made whether the hemolysis is immune-mediated or of non-immune origin. We report a case of hereditary spherocytosis in a 12-yr-old male child, in whom flow-assisted diagnosis was made. In this case, diagnosis was not determined because routine laboratory workups for hereditary spherocytosis yielded discrepant RESULTS: positive osmotic fragility test, positive direct antiglobulin test, and normal result in the red cell membrane protein sodium dodecyl succinimide polyacrylamide gel electrophoresis. However, all flow cytometry-based tests, such as osmotic fragility, direct antiglobulin, and eosin 5-maleimide binding test, yielded results compatible with hereditary spherocytosis. Additionally, in family study, the results of eosin 5-maleimide binding test suggested his disease being hereditary. In cases with diagnostic difficulties, flow cytometry may be used as an alternative tool, which can provide additional information in the differential diagnosis of hemolytic anemia with spherocytosis.
Anemia, Hemolytic/complications/*diagnosis ; Child ; Coombs' Test ; Diagnosis, Differential ; Eosine Yellowish-(YS)/analogs & derivatives/chemistry ; Erythrocytes/immunology/metabolism ; Flow Cytometry ; Humans ; Male ; Osmotic Fragility ; Spherocytosis, Hereditary/complications/*diagnosis

Astragalus heteropolysaccharides (AHPS) is obtained from the dried roots of Astragalus membranaceus (Fisch.) Bunge var. mongholious (Bunge) Hsiao. In the present study, we observed its effects on erythrocyte immune adherence function in mice with adjuvant-induced arthritis (AA). The mice were treated intragastrically with AHPS of 1 000, 500, and 250 mg x kg(-1) x d(-1) separately and treated with tripterygium glycosides (TG) of 60 mg x kg(-1) x d(-1) as positive control. The number of complement receptor type 1 (CR1) on erythrocyte, the concentration of circulating immune complex (CIC) in serum and the amount of immune complex (IC) deposition in synovium of knee joint were determined by flow cytometry, polyethylene glycol (PEG-6000) precipitation and ponceau S (P-S) staining and fluorescent immunohistochemistry respectively. The pathological change of knee joint was evaluated by histological section. The results showed that both AHPS and TG improved significantly the primary and secondary local or systemic symptoms of the mice with AA and reduced the synovium hyperplasia, inflammatory cell infiltrate, pannus and cartilage demolish of knee joint, and AHPS of 1 000, 500, and 250 mg x kg(-1) x d(-1) could significantly increase the number of CR1 on erythrocyte, improve the elimination of CIC in the peripheral blood and reduce the deposition of IC in joint synovium in a dose-dependent manner (P < 0.01 or P < 0.05). The results indicate that one of the therapeutic effective mechanisms of AHPS on mice with AA could be to increase gene expression of CR1 of mice with AA.
Animals ; Antigen-Antibody Complex ; blood ; metabolism ; Arthritis, Experimental ; metabolism ; pathology ; Astragalus Plant ; chemistry ; Dose-Response Relationship, Drug ; Erythrocytes ; immunology ; Knee Joint ; pathology ; Male ; Mice ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polysaccharides ; administration & dosage ; isolation & purification ; pharmacology ; Random Allocation ; Receptors, Complement ; blood ; Synovial Membrane ; immunology

Simultaneous drug-induced immune hemolytic anemia (DIIHA) caused by multiple drugs is rare. We report a case of a patient who developed DIIHA caused by 2 drugs. The patient's serum exhibited agglutination of ceftizoxime- or sulbactam-coated red blood cells (RBCs; via a drug-adsorption mechanism) and of uncoated RBCs in the presence of sulbactam (via an immune-complex mechanism). Although ceftizoxime is known to exhibit a positive reaction by an immune-complex method with or without reactivity with drug-coated RBCs, this patient's antibodies were reactive only against drug-coated RBCs. On the other hand, sulbactam, which is known to cause hemolytic anemia by nonimmunologic protein adsorption, exhibited positive reactions in tests with both drug-coated RBCs and in the presence of sulbactam. This is the first report of DIIHA due to a sulbactam-cefoperazone combination and the fourth report of DIIHA due to ceftizoxime. Owing to the patient's complicated laboratory results, DIIHA was suspected only at a late stage. We propose that for the prompt diagnosis of DIIHA, tests for all possible causative drugs should be conducted by 2 methods.
Anemia, Hemolytic/chemically induced/*diagnosis/immunology ; Anti-Bacterial Agents/*adverse effects ; Cefoperazone/*adverse effects ; Ceftizoxime/*adverse effects ; Erythrocytes/chemistry/metabolism ; Female ; Humans ; Middle Aged ; Sulbactam/*adverse effects