To examine the association between a multibiomarker disease activity (MBDA) score, CRP and clinic... more To examine the association between a multibiomarker disease activity (MBDA) score, CRP and clinical disease activity measures among RA patients with and without concomitant FM. In an observational cohort of patients with established RA, we performed a cross-sectional analysis comparing MBDA scores with CRP by rank correlation and cross-classification. MBDA scores, CRP and clinical measures of disease activity were compared between patients with RA alone and RA with concomitant FM (RA and FM) by univariate and multivariate analyses. CRP was ⩽1.0 mg/dl for 184 of 198 patients (93%). MBDA scores correlated with CRP (r = 0.755, P < 0.001), but were often discordant, being moderate or high for 19%, 55% and 87% of patients with CRP ⩽0.1, 0.1 to ⩽0.3, or 0.3 to ⩽1.0 mg/dl, respectively. Among patients with CRP ⩽1.0 mg/dl, swollen joint count (SJC) increased linearly across levels of MBDA score, both with (P = 0.021) and without (P = 0.004) adjustment for CRP, whereas CRP was not associa...
Objective. Rheumatoid arthritis (RA) clinical trials often exclude patients who have low C-reacti... more Objective. Rheumatoid arthritis (RA) clinical trials often exclude patients who have low C-reactive protein (CRP) levels, which slows enrollment into the trial. The purpose of this study was to determine whether high Multi-Biomarker Disease Activity (MBDA) scores (>44) in RA patients with low CRP levels (£10 mg/liter) could be used as a complement to CRP levels >10 mg/ liter to enhance patient recruitment without affecting clinical trial outcomes. Methods. We evaluated patients from the Swedish Pharmacotherapy (SWEFOT) trial, which did not include any selection criteria for CRP levels. Clinical ClinicalTrials.gov identifier: NCT00764725. Crescendo Bioscience, Inc., a wholly owned subsidiary of Myriad Genetics, Inc., performed the analyses of the MBDA scores at no cost to the investigators.
Objective. To evaluate the efficacy and safety of adalimumab plus methotrexate (MTX) administered... more Objective. To evaluate the efficacy and safety of adalimumab plus methotrexate (MTX) administered up to 4 years in patients with active, long-standing rheumatoid arthritis (RA). Methods. Patients responding inadequately to MTX who entered a 24-week, controlled study (ARMADA) with adalimumab plus MTX or placebo plus MTX and those who enrolled in a subsequent open-label extension were evaluated for efficacy and safety. Additional analyses were conducted for those patients allowed to adjust corticosteroid and/or MTX dosages during the extension. Results. Of 271 patients in the original ARMADA trial, 262 patients received at least one dose of adalimumab and were evaluated. At the time of analysis, 162 (62%) patients had remained in the study for a mean of 3.4 years on therapy. Withdrawals were for lack of efficacy (8%), adverse events (12%), and other reasons (18%). In 147 patients who had completed 4 years of therapy, efficacy improvements achieved at 6 months were maintained. At 4 years, 79%, 58%, and 39% had achieved ACR20/50/70; 43% achieved clinical remission (DAS28<2.6) and 22% had no physical function abnormalities (Health Assessment Questionnaire=0). Results were similar for 197 patients who were on therapy 2-4 years. Large percentages of patients were able to decrease dosages of corticosteroids (63%), MTX (42%), or both (12%), while sustaining efficacy. Serious adverse events during open-label therapy were comparable to those seen in the controlled phase. The rate of serious infections was 2.03 events per 100-patient years vs. 2.30 during the blinded period. Conclusions. Adalimumab plus MTX sustained clinical response and remission in RA patients during 4 years of observation. The safety profile observed during the first 6 months was similar to the profile after 4 years of follow-up. Patients were able to reduce MTX and/or corticosteroid dosages without adversely affecting long-term efficacy.
Introduction Comprehensive genomic profiling of solid tumors using circulating tumor DNA (ctDNA) ... more Introduction Comprehensive genomic profiling of solid tumors using circulating tumor DNA (ctDNA) has enabled the detection of all NCCN guideline-recommended somatic genomic classes of alterations from a single, non-invasive blood draw. However, current ctDNA tests still face two major challenges: the inability to reliably identify somatic variants at low mutant allele fraction (MAF), and inconsistency in how the tests have been validated. This study shows how the Single Molecule Sequencing (SMSEQ) platform addresses these challenges. The platform integrates innovative ctDNA extraction methodology, highly optimized library preparation and an error-based variant-calling algorithm to drastically improve sensitivity and specificity. The platform analyzes 5 classes of somatic variants: single nucleotide variants (SNVs), insertions and deletions (Indels), copy number variants (CNVs), fusions and microsatellite instability (MSI). Methods We analyzed a 73 gene panel covering NCCN recommende...
143 Background: The diagnostic confirmation of prostate cancer in patients with a PSA in the gray... more 143 Background: The diagnostic confirmation of prostate cancer in patients with a PSA in the gray zone (4–10 ng/ml) is controversial, often leading to unnecessary biopsies. The sensitivity of the PSA test at a 4 ng/mL cut-off can be as low as 21%. We introduce a new test for prostate cancer detection in PSA gray zone patients, with the potential to decrease the number of unnecessary prostate biopsies. Methods: A single-center, multi-year, IRB-approved, prospective, blinded clinical study was conducted in 200 high-risk subjects. 4 mL of blood was drawn and processed for CTC analysis using the CellMax biomimetic platform (CMx). The CMx CTC test uses a proprietary microfluidic biochip that accurately captures and enumerates CTCs with antibodies to EpCAM, CK18 and PSMA. All patients underwent routine screening including PSA and digital rectal exam (DRE). Gray zone and those diagnosed as ‘diseased’ based on PSA & DRE also underwent a biopsy. Multivariate generalized linear models incorpo...
e13549Background: Early detection is key to reducing cancer deaths. Despite available screening o... more e13549Background: Early detection is key to reducing cancer deaths. Despite available screening options for many cancers, patients are still diagnosed in advanced stages. A sensitive, affordable blood test could improve testing compliance, and detect cancer at earlier, more treatable stages. Here we present results from prospective, multi-center, IRB-approved studies for colorectal, prostate and breast cancer. Methods: 2 to 4 mL blood were processed through the CMx test platform which includes biomimetic chip, proprietary EpCAM antibody and an air foam release mechanism to increase sensitivity. Released CTCs were confirmed with tissue-specific antibodies CK20, PSMA and mammaglobin. The diagnosis and staging of cancers was confirmed by biopsy. For colorectal and prostate cancer, a scoring system was developed based on an algorithm that incorporated CTCs and age. For breast cancer, a preliminary scoring is being evaluated. Results: In colorectal cancer, 620 subjects were evaluated and the test had a sensiti...
Objective To evaluate the analytical performance of a 98-gene NGS panel designed to detect highly... more Objective To evaluate the analytical performance of a 98-gene NGS panel designed to detect highly- penetrant, rare pathogenic germline variants, strongly linked to predisposition of cancers in Taiwan and USA. Methods This 98-gene panel was developed following an extensive review of genes with a strong clinical and genetic linkage evidence for cancer predisposition. Probes covering the exon sequences from these 98 genes were validated using 19 samples obtained from the Platinum Genomes Project and the Genome In A Bottle Consortium. Analytical sensitivity and specificity for the detection of clinically important variants within multiple cancers types were demonstrated in 30 reference samples derived from patients with familial cancers. Additionally, two clinical cohorts of 1885 and 374 subjects from Taiwan and USA, respectively, were screened for detection of rare pathogenic germline variants in our CAP accredited laboratory in Taiwan, and CLIA certified laboratory in the USA. Results...
556 Background: Colorectal cancer (CRC) is among the most preventable cancers when precancerous l... more 556 Background: Colorectal cancer (CRC) is among the most preventable cancers when precancerous lesions are detected at an early stage. Current screening methods for CRC require bowel prep or stool-based testing that are inconvenient, resulting in low compliance. Stool based tests have limited sensitivity for the detection of precancerous lesions. We have conducted a prospective clinical study over a period of > 3 years to assess a novel assay to detect and enumerate circulating tumor cells (CTCs) in a blood sample for early CRC detection. Methods: A single-center, IRB-approved, prospective and blinded clinical study was conducted in 620 subjects including 438 with adenoma, polyps or stage I-IV CRC and 182 healthy controls. For each subject, 2mL peripheral whole blood collected through a routine blood draw was processed using the CellMax biomimetic platform (CMx). The CMx test is a proprietary microfluidic biochip that minimizes non-specific binding and accurately enumerates CTCs...
To analyze 2 indices composed of the 3 patient reported outcomes (PRO) in the American College of... more To analyze 2 indices composed of the 3 patient reported outcomes (PRO) in the American College of Rheumatology (ACR) Core Data Set--physical function, pain, and global estimate--without joint count or laboratory data, for capacities to distinguish active from control treatments in 4 pivotal clinical trials. Data from 4 clinical trials involving adalimumab, in combination with methotrexate or other disease-modifying antirheumatic drugs (DMARD) or as monotherapy, versus control treatment were made available to analyze properties of various indices. A categorical PRO-Index M was defined as "majority" improvement in 2 of the 3 PRO measures at 20%, 50%, and 70% levels; results were evaluated to analyze agreement with ACR20, ACR50, ACR70 responses and an "all Core Data Set measures" index based on 4 of the 7 measures having such levels of improvement. A continuous PRO-Index C was defined as the median or 2nd highest of 3 percentage differences from baseline to endpoint...
When studied by immunoelectrophoresis the factor B (BF)*F allele is a monomorphic protein, but by... more When studied by immunoelectrophoresis the factor B (BF)*F allele is a monomorphic protein, but by using isoelectric focusing (IEF), it turns out to be polymorphic. Two BF*F allelic subdivisions, BF*FA and BF*FB, were detected in samples from unrelated Spanish donors; *FA and *FB are in Hardy-Weinberg equilibrium, segregate as autosomal codominant alleles in families, and BF*FA (but not BF*FB) is found to be in linkage disequilibrium with B44 and DR7 and is within the A29, B44, Bw4, Cx, CFA31, DR7, DRw53, DQ2 haplotype. Furthermore, BF FA individuals have a higher BF serum concentration than the BF FB individuals. The subdivision of BF S observed with IEF was found to be due to nonspecific BF S cleavage by serum proteases in inadequately collected or aged samples. Thus, the subdivision of BF S is spurious and was not found in our sera. BF F1 and BF SO7 could not be further subdivided by IEF in our subjects. The BF F1 banding pattern was characterised by the presence of a cathodal band which corresponds to the Bb activation fragment. Finally, IEF combined with immunoblotting and monoclonal Ba and Bb antibodies may be used for accurately distinguishing BF phenotypes and doubtful bands.
Three novel restriction fragment length polymorphisms (RFLPs) have been identified using a pan-HL... more Three novel restriction fragment length polymorphisms (RFLPs) have been identified using a pan-HLA class I probe and the endonuclease SstI. This study, in conjunction with previously reported SstI RFLPs, now allows the identification of the HLA-crossreacting antigens Aw19 (A29/30/31/32/w33), A23/24 and A3/11 by specific hybridization patterns with a single enzyme/probe combination. Three of the corresponding polymorphic SstI restriction sites map within the HLA-A gene and generate two allelic RFLPs (5.06, 5.92 kb) and one single RFLP (5.92 kb) that show an absolute correlation with HLA-A23/24 and A29/32 crossreacting antigens, respectively. However, other SstI RFLPs (7.97, 9.4, 9.6, 9.8 and 13.34 kb), also linked to HLA-A crossreacting antigens, map outside the HLA-A gene and probably correspond to non-HLA-A,B,C class I genes in strong linkage disequilibrium with the HLA-A gene. These data show that HLA-A crossreacting antigens share more SstI RFLPs than neighboring non-HLA-A,B,C class I genes or pseudogenes; also, this has raised the possibility that some crossreacting HLA-A alloantisera might additionally recognize shared antigenic determinants in non-HLA-A,B,C proteins since the HLA-Aw19 crossreactivity cannot be fully explained by analyzing the HLA-A amino acid sequence.
Source and Description ofProbe: The probe JA8-4 BE0.6 is a 600 bp single-copy BamHI/EcoRI fragmen... more Source and Description ofProbe: The probe JA8-4 BE0.6 is a 600 bp single-copy BamHI/EcoRI fragment derived from a X phage clone. XJA8-I was obtained by screening a human chromosome jumping library (1) with the probe A8 (2). The sequence was cloned into the pUC19 plasmid at BamHI and EcoRI sites to yield the clone pJA8-l BEO.6 which was transformed into DH5a E. coli cells.
A simple and highly sensitive analytical isoelectric focusing (IEF) technique using immunofixatio... more A simple and highly sensitive analytical isoelectric focusing (IEF) technique using immunofixation with anti-C2 serum followed by silver staining has been developed in order to study simultaneously the structural polymorphism of both native C2 and C2 activation fragments (C2a MW 74 000 and C2b MW 34000). Structural C2 polymorphism of C2*B and C2"C allotypes (but not of C2"A1) was found to be associated with the C2a fragment, whereas C2b appears to display no polymorphism. Commercially available IEF gels and C2 antisera gave reproducible allotyping data and make this technique of general use for densitometric analysis of native and activated C2. In vivo C2 activation was studied through C2a/native C2 area ratios obtained by computerized densitometry. Significantly lower ratios were observed in healthy individuals than in patients with systemic lupus erythematosus (SLE), reflecting an abnormally high classical pathway activation of complement in SLE. This methodology may be of value for immunogenetic and functional studies of other complement components.
A T cell line termed DIL2 has been derived from an infant with a polyclonal T cell receptor (TCR)... more A T cell line termed DIL2 has been derived from an infant with a polyclonal T cell receptor (TCR)/CD3 cell surface expression defect. Indirect immunofluorescence showed that the expression of certain TCR/CD3 epitopes (like those detected by WT31 and BMA031 monoclonals) was strongly reduced (around five-fold) on DIL2, whereas other epitopes (like those detected by SP34 and Leu4) were only around two-fold lower than in normal T cell lines. Specific immunoprecipitates of surface-radioiodinated DIL2 cells contained TCR-alpha, TCR-beta, CD3-delta, CD3-epsilon and TCR-zeta chains, but lacked CD3-gamma. This structural TCR/CD3 variant was, however, capable of transducing certain activation signals, since normal proliferation and a low but significant calcium flux was observed in DIL2 cells after engagement with specific antibodies. Our data suggest that a functional TCR/CD3 complex can be expressed on the surface of T cells in the absence of CD3-gamma.
... Oscar G. Segurado xy, Paz Iglesias-Casarrubios l, Pablo Morales 1, Jorge Martinez-Laso 1, Juk... more ... Oscar G. Segurado xy, Paz Iglesias-Casarrubios l, Pablo Morales 1, Jorge Martinez-Laso 1, Jukka Partanen 2, R. Duncan Campbell 3, and ... Tissue Antigens 20: 123-140, 1982 Dunham, I., Sargent, CA, Trowsdale, J., and Campbell, RD: Molecular mapping of the human major ...
The combination of the HLA complement allorypes BFS, C2C, C4AQO (deleted gene) and C4B1, termed S... more The combination of the HLA complement allorypes BFS, C2C, C4AQO (deleted gene) and C4B1, termed SC01 complorype, usually present in the HLA-BS,DR3,DQw2 diabetogenic haplotype, has also been found in a novel <dow frequency» HLA-B49,DR4,DQwS haplorype associated with Spanish insulin-dependent diabetes mellitus (IDDM). Family studies of C4 antigenic determinants Rodgers/Chido and their specific C4d nucleotide sequences confirm that this novel haplotype bearing Chido-3,-6 is not due to a recent recombination from the common HLA-BS,DR3 haplotype bearing Chido 3,6; moreover, Chido analysis at the serological or DNA level is presently the only way to distinguish both SC01 complotypes, since BF, C2, steroid 21-hydroxylase and C4 genes do not reveal other differences by restriction fragment analysis. On the other hand, HLA-B49,SC01,DR4 is the first DR4bearing IDDM-susceptible haplorype with a deleted C4 gene described so far and the only DR4-bearing haplotype found in the Spanish population. This report further supports the fact that extended haplotypes with deleted (or «not duplicated») genes in the class III region contain IDDM-susceptibility more often than non-deleted (or «duplicated») haplotypes in the Spanish and other Mediterranean populations.
To examine the association between a multibiomarker disease activity (MBDA) score, CRP and clinic... more To examine the association between a multibiomarker disease activity (MBDA) score, CRP and clinical disease activity measures among RA patients with and without concomitant FM. In an observational cohort of patients with established RA, we performed a cross-sectional analysis comparing MBDA scores with CRP by rank correlation and cross-classification. MBDA scores, CRP and clinical measures of disease activity were compared between patients with RA alone and RA with concomitant FM (RA and FM) by univariate and multivariate analyses. CRP was ⩽1.0 mg/dl for 184 of 198 patients (93%). MBDA scores correlated with CRP (r = 0.755, P < 0.001), but were often discordant, being moderate or high for 19%, 55% and 87% of patients with CRP ⩽0.1, 0.1 to ⩽0.3, or 0.3 to ⩽1.0 mg/dl, respectively. Among patients with CRP ⩽1.0 mg/dl, swollen joint count (SJC) increased linearly across levels of MBDA score, both with (P = 0.021) and without (P = 0.004) adjustment for CRP, whereas CRP was not associa...
Objective. Rheumatoid arthritis (RA) clinical trials often exclude patients who have low C-reacti... more Objective. Rheumatoid arthritis (RA) clinical trials often exclude patients who have low C-reactive protein (CRP) levels, which slows enrollment into the trial. The purpose of this study was to determine whether high Multi-Biomarker Disease Activity (MBDA) scores (>44) in RA patients with low CRP levels (£10 mg/liter) could be used as a complement to CRP levels >10 mg/ liter to enhance patient recruitment without affecting clinical trial outcomes. Methods. We evaluated patients from the Swedish Pharmacotherapy (SWEFOT) trial, which did not include any selection criteria for CRP levels. Clinical ClinicalTrials.gov identifier: NCT00764725. Crescendo Bioscience, Inc., a wholly owned subsidiary of Myriad Genetics, Inc., performed the analyses of the MBDA scores at no cost to the investigators.
Objective. To evaluate the efficacy and safety of adalimumab plus methotrexate (MTX) administered... more Objective. To evaluate the efficacy and safety of adalimumab plus methotrexate (MTX) administered up to 4 years in patients with active, long-standing rheumatoid arthritis (RA). Methods. Patients responding inadequately to MTX who entered a 24-week, controlled study (ARMADA) with adalimumab plus MTX or placebo plus MTX and those who enrolled in a subsequent open-label extension were evaluated for efficacy and safety. Additional analyses were conducted for those patients allowed to adjust corticosteroid and/or MTX dosages during the extension. Results. Of 271 patients in the original ARMADA trial, 262 patients received at least one dose of adalimumab and were evaluated. At the time of analysis, 162 (62%) patients had remained in the study for a mean of 3.4 years on therapy. Withdrawals were for lack of efficacy (8%), adverse events (12%), and other reasons (18%). In 147 patients who had completed 4 years of therapy, efficacy improvements achieved at 6 months were maintained. At 4 years, 79%, 58%, and 39% had achieved ACR20/50/70; 43% achieved clinical remission (DAS28<2.6) and 22% had no physical function abnormalities (Health Assessment Questionnaire=0). Results were similar for 197 patients who were on therapy 2-4 years. Large percentages of patients were able to decrease dosages of corticosteroids (63%), MTX (42%), or both (12%), while sustaining efficacy. Serious adverse events during open-label therapy were comparable to those seen in the controlled phase. The rate of serious infections was 2.03 events per 100-patient years vs. 2.30 during the blinded period. Conclusions. Adalimumab plus MTX sustained clinical response and remission in RA patients during 4 years of observation. The safety profile observed during the first 6 months was similar to the profile after 4 years of follow-up. Patients were able to reduce MTX and/or corticosteroid dosages without adversely affecting long-term efficacy.
Introduction Comprehensive genomic profiling of solid tumors using circulating tumor DNA (ctDNA) ... more Introduction Comprehensive genomic profiling of solid tumors using circulating tumor DNA (ctDNA) has enabled the detection of all NCCN guideline-recommended somatic genomic classes of alterations from a single, non-invasive blood draw. However, current ctDNA tests still face two major challenges: the inability to reliably identify somatic variants at low mutant allele fraction (MAF), and inconsistency in how the tests have been validated. This study shows how the Single Molecule Sequencing (SMSEQ) platform addresses these challenges. The platform integrates innovative ctDNA extraction methodology, highly optimized library preparation and an error-based variant-calling algorithm to drastically improve sensitivity and specificity. The platform analyzes 5 classes of somatic variants: single nucleotide variants (SNVs), insertions and deletions (Indels), copy number variants (CNVs), fusions and microsatellite instability (MSI). Methods We analyzed a 73 gene panel covering NCCN recommende...
143 Background: The diagnostic confirmation of prostate cancer in patients with a PSA in the gray... more 143 Background: The diagnostic confirmation of prostate cancer in patients with a PSA in the gray zone (4–10 ng/ml) is controversial, often leading to unnecessary biopsies. The sensitivity of the PSA test at a 4 ng/mL cut-off can be as low as 21%. We introduce a new test for prostate cancer detection in PSA gray zone patients, with the potential to decrease the number of unnecessary prostate biopsies. Methods: A single-center, multi-year, IRB-approved, prospective, blinded clinical study was conducted in 200 high-risk subjects. 4 mL of blood was drawn and processed for CTC analysis using the CellMax biomimetic platform (CMx). The CMx CTC test uses a proprietary microfluidic biochip that accurately captures and enumerates CTCs with antibodies to EpCAM, CK18 and PSMA. All patients underwent routine screening including PSA and digital rectal exam (DRE). Gray zone and those diagnosed as ‘diseased’ based on PSA & DRE also underwent a biopsy. Multivariate generalized linear models incorpo...
e13549Background: Early detection is key to reducing cancer deaths. Despite available screening o... more e13549Background: Early detection is key to reducing cancer deaths. Despite available screening options for many cancers, patients are still diagnosed in advanced stages. A sensitive, affordable blood test could improve testing compliance, and detect cancer at earlier, more treatable stages. Here we present results from prospective, multi-center, IRB-approved studies for colorectal, prostate and breast cancer. Methods: 2 to 4 mL blood were processed through the CMx test platform which includes biomimetic chip, proprietary EpCAM antibody and an air foam release mechanism to increase sensitivity. Released CTCs were confirmed with tissue-specific antibodies CK20, PSMA and mammaglobin. The diagnosis and staging of cancers was confirmed by biopsy. For colorectal and prostate cancer, a scoring system was developed based on an algorithm that incorporated CTCs and age. For breast cancer, a preliminary scoring is being evaluated. Results: In colorectal cancer, 620 subjects were evaluated and the test had a sensiti...
Objective To evaluate the analytical performance of a 98-gene NGS panel designed to detect highly... more Objective To evaluate the analytical performance of a 98-gene NGS panel designed to detect highly- penetrant, rare pathogenic germline variants, strongly linked to predisposition of cancers in Taiwan and USA. Methods This 98-gene panel was developed following an extensive review of genes with a strong clinical and genetic linkage evidence for cancer predisposition. Probes covering the exon sequences from these 98 genes were validated using 19 samples obtained from the Platinum Genomes Project and the Genome In A Bottle Consortium. Analytical sensitivity and specificity for the detection of clinically important variants within multiple cancers types were demonstrated in 30 reference samples derived from patients with familial cancers. Additionally, two clinical cohorts of 1885 and 374 subjects from Taiwan and USA, respectively, were screened for detection of rare pathogenic germline variants in our CAP accredited laboratory in Taiwan, and CLIA certified laboratory in the USA. Results...
556 Background: Colorectal cancer (CRC) is among the most preventable cancers when precancerous l... more 556 Background: Colorectal cancer (CRC) is among the most preventable cancers when precancerous lesions are detected at an early stage. Current screening methods for CRC require bowel prep or stool-based testing that are inconvenient, resulting in low compliance. Stool based tests have limited sensitivity for the detection of precancerous lesions. We have conducted a prospective clinical study over a period of > 3 years to assess a novel assay to detect and enumerate circulating tumor cells (CTCs) in a blood sample for early CRC detection. Methods: A single-center, IRB-approved, prospective and blinded clinical study was conducted in 620 subjects including 438 with adenoma, polyps or stage I-IV CRC and 182 healthy controls. For each subject, 2mL peripheral whole blood collected through a routine blood draw was processed using the CellMax biomimetic platform (CMx). The CMx test is a proprietary microfluidic biochip that minimizes non-specific binding and accurately enumerates CTCs...
To analyze 2 indices composed of the 3 patient reported outcomes (PRO) in the American College of... more To analyze 2 indices composed of the 3 patient reported outcomes (PRO) in the American College of Rheumatology (ACR) Core Data Set--physical function, pain, and global estimate--without joint count or laboratory data, for capacities to distinguish active from control treatments in 4 pivotal clinical trials. Data from 4 clinical trials involving adalimumab, in combination with methotrexate or other disease-modifying antirheumatic drugs (DMARD) or as monotherapy, versus control treatment were made available to analyze properties of various indices. A categorical PRO-Index M was defined as "majority" improvement in 2 of the 3 PRO measures at 20%, 50%, and 70% levels; results were evaluated to analyze agreement with ACR20, ACR50, ACR70 responses and an "all Core Data Set measures" index based on 4 of the 7 measures having such levels of improvement. A continuous PRO-Index C was defined as the median or 2nd highest of 3 percentage differences from baseline to endpoint...
When studied by immunoelectrophoresis the factor B (BF)*F allele is a monomorphic protein, but by... more When studied by immunoelectrophoresis the factor B (BF)*F allele is a monomorphic protein, but by using isoelectric focusing (IEF), it turns out to be polymorphic. Two BF*F allelic subdivisions, BF*FA and BF*FB, were detected in samples from unrelated Spanish donors; *FA and *FB are in Hardy-Weinberg equilibrium, segregate as autosomal codominant alleles in families, and BF*FA (but not BF*FB) is found to be in linkage disequilibrium with B44 and DR7 and is within the A29, B44, Bw4, Cx, CFA31, DR7, DRw53, DQ2 haplotype. Furthermore, BF FA individuals have a higher BF serum concentration than the BF FB individuals. The subdivision of BF S observed with IEF was found to be due to nonspecific BF S cleavage by serum proteases in inadequately collected or aged samples. Thus, the subdivision of BF S is spurious and was not found in our sera. BF F1 and BF SO7 could not be further subdivided by IEF in our subjects. The BF F1 banding pattern was characterised by the presence of a cathodal band which corresponds to the Bb activation fragment. Finally, IEF combined with immunoblotting and monoclonal Ba and Bb antibodies may be used for accurately distinguishing BF phenotypes and doubtful bands.
Three novel restriction fragment length polymorphisms (RFLPs) have been identified using a pan-HL... more Three novel restriction fragment length polymorphisms (RFLPs) have been identified using a pan-HLA class I probe and the endonuclease SstI. This study, in conjunction with previously reported SstI RFLPs, now allows the identification of the HLA-crossreacting antigens Aw19 (A29/30/31/32/w33), A23/24 and A3/11 by specific hybridization patterns with a single enzyme/probe combination. Three of the corresponding polymorphic SstI restriction sites map within the HLA-A gene and generate two allelic RFLPs (5.06, 5.92 kb) and one single RFLP (5.92 kb) that show an absolute correlation with HLA-A23/24 and A29/32 crossreacting antigens, respectively. However, other SstI RFLPs (7.97, 9.4, 9.6, 9.8 and 13.34 kb), also linked to HLA-A crossreacting antigens, map outside the HLA-A gene and probably correspond to non-HLA-A,B,C class I genes in strong linkage disequilibrium with the HLA-A gene. These data show that HLA-A crossreacting antigens share more SstI RFLPs than neighboring non-HLA-A,B,C class I genes or pseudogenes; also, this has raised the possibility that some crossreacting HLA-A alloantisera might additionally recognize shared antigenic determinants in non-HLA-A,B,C proteins since the HLA-Aw19 crossreactivity cannot be fully explained by analyzing the HLA-A amino acid sequence.
Source and Description ofProbe: The probe JA8-4 BE0.6 is a 600 bp single-copy BamHI/EcoRI fragmen... more Source and Description ofProbe: The probe JA8-4 BE0.6 is a 600 bp single-copy BamHI/EcoRI fragment derived from a X phage clone. XJA8-I was obtained by screening a human chromosome jumping library (1) with the probe A8 (2). The sequence was cloned into the pUC19 plasmid at BamHI and EcoRI sites to yield the clone pJA8-l BEO.6 which was transformed into DH5a E. coli cells.
A simple and highly sensitive analytical isoelectric focusing (IEF) technique using immunofixatio... more A simple and highly sensitive analytical isoelectric focusing (IEF) technique using immunofixation with anti-C2 serum followed by silver staining has been developed in order to study simultaneously the structural polymorphism of both native C2 and C2 activation fragments (C2a MW 74 000 and C2b MW 34000). Structural C2 polymorphism of C2*B and C2"C allotypes (but not of C2"A1) was found to be associated with the C2a fragment, whereas C2b appears to display no polymorphism. Commercially available IEF gels and C2 antisera gave reproducible allotyping data and make this technique of general use for densitometric analysis of native and activated C2. In vivo C2 activation was studied through C2a/native C2 area ratios obtained by computerized densitometry. Significantly lower ratios were observed in healthy individuals than in patients with systemic lupus erythematosus (SLE), reflecting an abnormally high classical pathway activation of complement in SLE. This methodology may be of value for immunogenetic and functional studies of other complement components.
A T cell line termed DIL2 has been derived from an infant with a polyclonal T cell receptor (TCR)... more A T cell line termed DIL2 has been derived from an infant with a polyclonal T cell receptor (TCR)/CD3 cell surface expression defect. Indirect immunofluorescence showed that the expression of certain TCR/CD3 epitopes (like those detected by WT31 and BMA031 monoclonals) was strongly reduced (around five-fold) on DIL2, whereas other epitopes (like those detected by SP34 and Leu4) were only around two-fold lower than in normal T cell lines. Specific immunoprecipitates of surface-radioiodinated DIL2 cells contained TCR-alpha, TCR-beta, CD3-delta, CD3-epsilon and TCR-zeta chains, but lacked CD3-gamma. This structural TCR/CD3 variant was, however, capable of transducing certain activation signals, since normal proliferation and a low but significant calcium flux was observed in DIL2 cells after engagement with specific antibodies. Our data suggest that a functional TCR/CD3 complex can be expressed on the surface of T cells in the absence of CD3-gamma.
... Oscar G. Segurado xy, Paz Iglesias-Casarrubios l, Pablo Morales 1, Jorge Martinez-Laso 1, Juk... more ... Oscar G. Segurado xy, Paz Iglesias-Casarrubios l, Pablo Morales 1, Jorge Martinez-Laso 1, Jukka Partanen 2, R. Duncan Campbell 3, and ... Tissue Antigens 20: 123-140, 1982 Dunham, I., Sargent, CA, Trowsdale, J., and Campbell, RD: Molecular mapping of the human major ...
The combination of the HLA complement allorypes BFS, C2C, C4AQO (deleted gene) and C4B1, termed S... more The combination of the HLA complement allorypes BFS, C2C, C4AQO (deleted gene) and C4B1, termed SC01 complorype, usually present in the HLA-BS,DR3,DQw2 diabetogenic haplotype, has also been found in a novel <dow frequency» HLA-B49,DR4,DQwS haplorype associated with Spanish insulin-dependent diabetes mellitus (IDDM). Family studies of C4 antigenic determinants Rodgers/Chido and their specific C4d nucleotide sequences confirm that this novel haplotype bearing Chido-3,-6 is not due to a recent recombination from the common HLA-BS,DR3 haplotype bearing Chido 3,6; moreover, Chido analysis at the serological or DNA level is presently the only way to distinguish both SC01 complotypes, since BF, C2, steroid 21-hydroxylase and C4 genes do not reveal other differences by restriction fragment analysis. On the other hand, HLA-B49,SC01,DR4 is the first DR4bearing IDDM-susceptible haplorype with a deleted C4 gene described so far and the only DR4-bearing haplotype found in the Spanish population. This report further supports the fact that extended haplotypes with deleted (or «not duplicated») genes in the class III region contain IDDM-susceptibility more often than non-deleted (or «duplicated») haplotypes in the Spanish and other Mediterranean populations.
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