Sensitized patients have a reduced chance of receiving a crossmath-negative organ, because of the presence of antibodies against a variety of human leukocyte antigen (HLA). There have been much attention and understanding on renal transplantation in sensitized patients, which led to attempts to remove alloantibodies and successful transplantation. Therefore, a positive crossmatch test and the presence of donor-specific anti-HLA antibodies are no longer a contraindication to renal transplantation. These desensitization protocols include intravenous immunoglobulin (IVIG), plasmapheresis (PP), and rituximab. IVIG therapy is performed in two forms, high dose IVIG and low dose IVIG in combination with PP. Rituximab is usually utilized in combination with IVIG therapy. Here we summarized the characterisitics of these strategies.
Antibodies ; Antibodies, Monoclonal, Murine-Derived ; Humans ; Immunoglobulins ; Immunoglobulins, Intravenous ; Isoantibodies ; Kidney Transplantation ; Leukocytes ; Plasmapheresis ; Rituximab ; Transplants

OBJECTIVE: To gain insights into structural characteristics of immunoglobulin kappa chain repertoire expressed in the inflammed synovium of rheumatoid arthritis (RA), we analyzed V kappa transcirpts expressed in the synovium of a patient with longstanding RA and compared to those expressed in the PBLs of RA and normal controls. METHODS: RT-PCR was done to amplify kappa chain transcripts from RA synovial lymphocytes and the cDNA sequences were compared to those from PBL of RA patient or normal control. RESULTS: Kappa chain repertoire from RA patient's synovial lymphocytes or PBL revealed increased somatic mutation and unusually long complementarity determining region (CDR) 3 compared to normal control. CONCLUSIONS: These changes in kappa chain repertoire in RA patient are suggesting that the antibody repertoire expressed in the synovium or PBL is unique and may be related with systemic dysregulation of B cell development
Arthritis, Rheumatoid ; Autoimmune Diseases* ; Complementarity Determining Regions ; DNA, Complementary ; Genes, Immunoglobulin* ; Humans ; Immunoglobulin kappa-Chains ; Immunoglobulins* ; Lymphocytes ; Synovial Membrane*

Phage display technology provides a powerful tool to screen a library for a binding molecule via an enrichment process. It has been adopted as a critical technology in the development of therapeutic antibodies. However, a major drawback of phage display technology is that because the degree of the enrichment cannot be controlled during the bio-panning process, it frequently results in a limited number of clones. In this study, we applied next-generation sequencing (NGS) to screen clones from a library and determine whether a greater number of clones can be identified using NGS than using conventional methods. Three chicken immune single-chain variable fragment (scFv) libraries were subjected to bio-panning on prostate-specific antigen (PSA). Phagemid DNA prepared from the original libraries as well as from the Escherichia coli pool after each round of bio-panning was analyzed using NGS, and the heavy chain complementarity-determining region 3 (HCDR3) sequences of the scFv clones were determined. Subsequently, through two-step linker PCR and cloning, the entire scFv gene was retrieved and analyzed for its reactivity to PSA in a phage enzyme immunoassay. After four rounds of bio-panning, the conventional colony screening method was performed for comparison. The scFv clones retrieved from NGS analysis included all clones identified by the conventional colony screening method as well as many additional clones. The enrichment of the HCDR3 sequence throughout the bio-panning process was a positive predictive factor for the selection of PSA-reactive scFv clones.
Antibodies ; Bacteriophages ; Chickens ; Clone Cells* ; Cloning, Organism ; Complementarity Determining Regions ; DNA ; Escherichia coli ; Immunoenzyme Techniques ; Mass Screening ; Methods ; Polymerase Chain Reaction ; Prostate-Specific Antigen ; Single-Chain Antibodies

PURPOSE: Antibody mediated rejection (AMR), although less common than acute cellular rejection (ACR), may be recalcitrant to conventional rescue therapy. AMR is caused by de novo B-cell mediated production of immunoglobulin G antibody (IgG) targeted against specific allograft antigen in a presensitized recipient. Rituximab is a chimeric murine- human anti-CD20 monoclonal antibody which targets CD-20 positive B-cells for elimination. Rituximab has been described to improve allograft salvage for refractory AMR. METHODS: From January 2002 to May 2004, 11 patients were diagnosed with AMR. The first 5 patients (non-rituximab group: NRG) were treated with high dose steroids, plasmapheresis followed by IVIG (500 mg/kg/dose) in addition to OKT3 and/or rabbit antithymocyte globulin. The latter 6 patients (rituximab group: RG) were given Rituximab (375 mg/m2) with IVIG following plasmapheresis. All patients had biopsy proven AMR. RESULTS: Four patients received allografts from living donors and one patient from cadaveric donor in NRG. Each three patients received allografts from living or cadaveric donors in RG. One patient of RG had a positive anti-HLA B-cell crossmatch by CDC (complement dependent cytotoxicity). The anti-donor antibody was reduced to zero with negative CDC and flowcytometry through a desensitization protocol prior to transplantation. The time to diagnosis of AMR in both groups were 17.8+/-18.17 days (NRG); 11+/-2.5 days (RG). ACR was identified in conjunction with AMR in 2 (40%: NRG), 4 patients (66.7%: RG), respectively. All patients had biopsies with classic features of AMR on light microscopy, including C4d staining. Three (50%) patients of RG had positive post-transplantation CDC and donor-specific antibody (DSA) identified. Mean serum creatinine (SCr) upon diagnosis of AMR were 4.3+/-1.71 mg/dL (NRG); 5.77+/-2.65 mg/dL (RG). The rescue rate of RG was superior than NRG (83% vs. 40%, P>0.05). The time to rescue from AMR in both groups were 40.5 +/-28.99 days (NRG); 48+/-54.67 days (RG). Mean SCr of the rescued patients were 1.65+/-0.07 mg/dL (NRG); 2.2+/-1.4 (RG) with median follow up of 120 days (range 33~319 days). Allograft nephrectomies were performed in 3 patients of NRG. CONCLUSION: Rescue therapy with Rituximab improves allograft salvage after AMR and should be considered early in the treatment of biopsy proven AMR.
Allografts ; Antilymphocyte Serum ; B-Lymphocytes ; Biopsy ; Cadaver ; Centers for Disease Control and Prevention (U.S.) ; Creatinine ; Diagnosis ; Follow-Up Studies ; Humans ; Immunoglobulin G ; Immunoglobulins, Intravenous ; Kidney Transplantation* ; Kidney* ; Living Donors ; Microscopy ; Muromonab-CD3 ; Nephrectomy ; Plasmapheresis ; Rituximab ; Steroids ; Tissue Donors

BACKGROUND: This study characterized clonal IG heavy V-D-J (IGH) gene rearrangements in South Indian patients with precursor B-cell acute lymphoblastic leukemia (precursor B-ALL) and identified age-related predominance in VDJ rearrangements. METHODS: IGH rearrangements were studied in 50 precursor B-ALL cases (common ALL=37, pre-B ALL=10, pro-B ALL=3) by polymerase chain reaction (PCR) heteroduplex analysis. Twenty randomly selected clonal IGH rearrangement sequences were analyzed using the IMGT/V-QUEST tool. RESULTS: Clonal IGH rearrangements were detected in 41 (82%) precursor B-ALL cases. Among the IGHV1-IGHV7 subgroups, IGHV3 was used in 25 (50%) cases. Among the IGHD1-IGHD7 genes, IGHD2 and IGHD3 were used in 8 (40%) and 5 (25%) clones, respectively. Among the IGHJ1-IGHJ6 genes, IGHJ6 and IGHJ4 were used in 9 (45%) and 6 (30%) clones, respectively. In 6 out of 20 (30%) IGH rearranged sequences, CDR3 was in frame whereas 14 (70%) had rearranged sequences and CDR3 was out of frame. A somatic mutation in Vmut/Dmut/Jmut was detected in 14 of 20 IGH sequences. On average, Vmut/Dmut/Jmut were detected in 0.1 nt, 1.1 nt, and 0.2 nt, respectively. CONCLUSION: The IGHV3 gene was frequently used whereas lower frequencies of IGHV5 and IGHV6 and a higher frequency of IGHV4 were detected in children compared with young adults. The IGHD2 and IGHD3 genes were over-represented, and the IGHJ6 gene was predominantly used in precursor-B-ALL. However, the IGH gene rearrangements in precursor-B-ALL did not show any significant age-associated genotype pattern attributed to our population.
Child ; Clone Cells ; Complementarity Determining Regions* ; Gene Rearrangement* ; Genotype ; Heteroduplex Analysis ; Humans ; Immunoglobulins* ; Polymerase Chain Reaction ; Precursor Cell Lymphoblastic Leukemia-Lymphoma* ; Precursor Cells, B-Lymphoid* ; Young Adult

Intravenous immunoglobulin (IVIG) infusion is an effective therapy for acute Kawasaki disease (KD). Nonetheless, approximately 10 percent to 20 percent of patients have persistent or recrudescent fever despite IVIG treatment, leading to a higher risk for coronary artery aneurysms (CAA). This unresponsiveness may pose a challenge to the clinicians. Tumor necrosis factor-alpha levels are elevated in the acute phase of the disease, especially in patients who develop CAA. We report a 10-month-old male with KD who failed to respond to multiple doses of IVIG and methylprednisolone and who then was treated with infliximab (5 mg/kg single dose). After infliximab treatment, he became afebrile with normalization of inflammatory markers and no further progression of CAA.
Aneurysm ; Coronary Vessels ; Fever ; Humans ; Immunoglobulins ; Immunoglobulins, Intravenous ; Infant ; Male ; Methylprednisolone ; Mucocutaneous Lymph Node Syndrome* ; Tumor Necrosis Factor-alpha ; Infliximab

Despite the recent advances in immunosuppression, antibody mediated acute rejection is associated with a poor prognosis. Therapeutic interventions such as polyclonal antibodies, rituximab, and plasmapheresis have been used for the treatment of antibody mediated rejection. However, these treatments are associated with a serious infectious complication. Recently, Intravenous immunoglobulin (IVIG) are known to have powerful immunomodulatory effects on inflammatory and infectious disease. We successfully rescued two patients who had concomitant antibody mediated rejection and BK virus infection. IVIG could be a preferable choice for treatment of humoral rejection in the presence of infection.
Antibodies ; BK Virus* ; Communicable Diseases ; Humans ; Immunoglobulins ; Immunoglobulins, Intravenous* ; Immunosuppression ; Kidney Transplantation ; Plasmapheresis ; Prognosis ; Rituximab ; Transplantation*

Group B streptococcus (GBS) infection is a leading cause of sepsis and meningitis among infants, and is associated with high rates of morbidity and mortality in many countries. Protection against GBS typically involves antibody-mediated opsonization by phagocytes and complement components. The present study evaluated serotype-specific functional antibodies to GBS among Korean infants and in intravenous immunoglobulin (IVIG) products. An opsonophagocytic killing assay (OPA) was used to calculate the opsonization indices (OIs) of functional antibodies to serotypes Ia, Ib, and III in 19 IVIG products from 5 international manufacturers and among 98 Korean infants (age: 0–11 months). The GBS Ia, Ib, and III serotypes were selected because they are included in a trivalent GBS vaccine formulation that is being developed. The OI values for the IVIG products were 635–5,706 (serotype Ia), 488–1,421 (serotype Ib), and 962–3,315 (serotype III), and none of the IVIG lots exhibited undetectable OI values (< 4). The geometric mean OI values were similar for all 3 serotypes when we compared the Korean manufacturers. The seropositive rate among infants was significantly lower for serotype Ia (18.4%), compared to serotype Ib and serotype III (both, 38.8%). Infant age of ≥ 3 months was positively correlated with the seropositive rates for each serotype. Therefore, only a limited proportion of infants exhibited protective immunity against serotype Ia, Ib, and III GBS infections. IVIG products that exhibit high antibody titers may be a useful therapeutic or preventive measure for infants. Further studies are needed to evaluate additional serotypes and age groups.
Antibodies* ; Complement System Proteins ; Homicide ; Humans ; Immunoglobulins* ; Immunoglobulins, Intravenous ; Infant* ; Meningitis ; Mortality ; Opsonin Proteins ; Phagocytes ; Sepsis ; Serogroup* ; Streptococcus agalactiae ; Streptococcus*

The third complementarity determining region (CDR3) of the immunoglobulin (Ig) kappa () chain is known to be located at the center of antigen binding groove and critical for antibody specificity. Ig chain has been characterized by limited junctional diversity due to the absence of N-region addition resulting in relative conservation of CDR3 lengths with 9 or 10 amino acids. CDR3 region of 11 amino acids is only possible with N-region addition. Recently, x transcripts with 11 amino acids CDR3 was found to be expressed in normal individuals, and in autoimrnune disease such as rheumatoid arthritis, the fraction of 11 amino acids CDR3 of humkv325-derived chains was overexpressed compared to conventional adult peripheral B cells. However, the significance of this bias is difficult to interpret without a clear understanding of normal repertoire of CDR3 length during development. The purpose of this study is to determine whether developmental regulation of CDR3 amino acids codon lengths exists in chains expressed in the fetal liver, cord blood, and adult peripheral blood lymphocytes (PBL). Lymphocytes were seperated from fetal liver, cord blood and adult PBL and cDNA was generated from extracted mRNA. PCR-based CDR3 finger- printing assay was performed with VI-IV family specific primers. CDR3 length diversity of Ig x chain increases as the development proceeds. The length diversity most frequently occured in Vlll family derived transcripts including 11 amino acids CDR3. transcripts with 11 amino acids CDR3 were consitently expressed in both fetal and adult Ig repertoire. These results support the hypothesis that v chain CDR3 length is developmentally regulated and implicates the diversity of antigen-antibody specificity generation.
Adult ; Amino Acids ; Antibody Specificity ; Arthritis, Rheumatoid ; B-Lymphocytes ; Bias (Epidemiology) ; Codon ; Complementarity Determining Regions ; DNA, Complementary ; Fetal Blood ; Humans* ; Immunoglobulin kappa-Chains* ; Immunoglobulins* ; Liver ; Lymphocytes ; RNA, Messenger ; Sensitivity and Specificity

A high degree of sensitization to human leukocyte antigen requires more intensive induction therapy; however, this increases vulnerability to opportunistic infections following kidney transplantation. Although recent studies have suggested that combined induction therapy with antithymocyte globulin and rituximab would be more effective in highly sensitized kidney recipients, we experienced a case of near-fatal invasive pulmonary aspergillosis 2 months after combined induction and early rejection therapy for graft dysfunction. Fortunately, the patient recovered with intensive antifungal treatment and lung lobectomy for a necrotic cavity. Antifungal prophylaxis should be considered in cases undergoing intensive induction therapy.
Antilymphocyte Serum* ; Humans ; Immunoglobulins* ; Invasive Pulmonary Aspergillosis* ; Kidney Transplantation* ; Kidney* ; Leukocytes ; Lung ; Opportunistic Infections ; Plasmapheresis* ; Rituximab* ; Transplants