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Removal of Coagulation Factors by the Gamunex®-C Purification Process
AB10 Abstracts
SATURDAY
32
Cytokine Profile and Clinical Correlates in Immune Deficient (HIV-infected) Infants with Severe (hypoxic) Pneumonia Robin J. Green, MD, PhD, FAAAAI1, Refiloe Masekela, MD, Cert Pulm Paed1, Dankwart Wittenberg, MD, Cert Pulm Paed1, Alta Terblanche, Cert Gastro Paed1, Paul Rheeder, PhD2, Piet Becker, PhD3, Ronald Anderson, PhD4; 1Department of Paediatrics and Child Health, University of Pretoria, South Africa, 2Division of Clinical Epidemiology, School of Health Systems and Public Health, University of Pretoria, Pretoria, South Africa, 3 Medical Research Council, South Africa, 4Medical Research Council Unit for Inflammation and Immunity, Department of Immunology, University of Pretoria and Tshwane Academic Division of National Health Laboratory Service, South Africa. RATIONALE: Severe pneumonia in infants who are HIV-infected is a common problem in many parts of the developing world, especially subSaharan Africa. What has been missing from previous studies of severe pneumonia in HIV-infected infants, however, is a description of the host inflammatory response and cytokine/chemokine profile that accompanies this disease. To describe the cytokine profiles associated with severe hypoxic pneumonia in HIV-infected infants. METHODS: In a cohort of HIV-infected children diagnosed clinically with severe hypoxic pneumonia, paired serum and sputum cytokines were tested. A control group of HIV-infected children with bronchiectasis contributed matching controls. RESULTS: A total of 100 infants (mean age 2.8 months) with a clinical diagnosis of severe hypoxic pneumonia were included in this study. IP-10 was markedly elevated in both sputum (mean 560.77pg/ml) and serum (mean 9091.14pg/ml), while IP-10 was elevated in serum (mean 39.55 pg/ml), with both these cytokines being significantly higher than in stable children with HIV-related bronchiectasis. CONCLUSIONS: This study of HIV-infected infants with severe hypoxic pneumonia suggests that IL-10 and IP-10 are associated with more severe lung disease. However, further investigation of this association is required.
33
Atheroprotection Conferred by Immunization with 15-Mer Peptide Fragments From ApoB100 Kevin Tse, MD1, Harley Tse, PhD2, Michael Shaw2, Alessandro Sette, Biol. Sci.3, Klaus Ley, MD3; 1UCSD, 2Wayne State University, 3 La Jolla Institute for Allergy and Immunology, La Jolla, CA. RATIONALE: Native LDL vaccinations of atherosclerotic-prone mice (ApoE-/-) have yielded 50% athero-reduction when administered repeatedly at regular intervals over several months. Akin to allergy immunotherapy, the presumed mechanism of this atheroprotection is thought to be due to induction of tolerance to the suspected autoantigen, oxidized LDL. Here, we report the discovery of a novel peptide found in murine ApoB100 (the lipoprotein portion of LDL) and demonstrate its ability to confer atheroprotection. METHODS: 15-mer peptide sequences spanning ApoB100 were synthesized and screened for their ability to bind tightly to MHC class II and cause T-cell proliferation after primary immunization. The candidate peptide (CP3) was used to vaccinate 5 female ApoE-/- using 50 micrograms of CP3/CFA subcutaneously in the inguinal area at 8 wks of age. Western diet was started at 10 wks of age. Repeated boosters with 25 micrograms of CP3/IFA were administered intraperitoneally at age 12, 16, 20 and 22 weeks. Mice were sacrificed at age 23 weeks and organs were harvested for analysis. PBS and irrelevant peptide were used as controls. RESULTS: CP3 treated mice showed >50% reduction in lesion size by aortic pinning when compared to PBS (p50.003) and irrelevant peptide controls (p50.004). No differences were observed in the percentage of IFNg, IL-2, IL-10, IL-17A, or FoxP3 positive cells in spleen or lymph nodes by intracellular FACS staining. CONCLUSIONS: CP3 is a novel peptide with T-cell specificity which affords atheroprotection to a similar degree as native LDL when used in a vaccination scheme.
J ALLERGY CLIN IMMUNOL FEBRUARY 2013
34
Removal of Coagulation Factors by the GamunexÒ-C Purification Process Melanie Williams, PhD, Jeff Willey, Katherine Tull, Glenn Stevenson, Mona Parks, LaToya McElrath, Luda Golovko, Catherine Russ, John Griffin, Pete Vandeberg; Grifols, Inc., Raleigh, NC. RATIONALE: GamunexÒ-C is a highly purified human plasma-derived intravenous immunoglobulin (IVIG). Coagulation factors are natural components of the starting plasma used for IVIG manufacture and are removed during purification. Residual coagulation factor activity has been associated with an increased incidence of thromboembolic adverse events with a specific immunoglobulin product. The purpose of this study was to characterize the removal of coagulation factors during the manufacture of GamunexÒ-C. METHODS: Relevant fractionation and purification process intermediates from 30 GamunexÒ-C batches were collected and analyzed for specific coagulation factors and procoagulant activity. The thrombin generation test (TGT) and the non-activated partial thromboplastin time (naPTT) are global coagulation methods that detect procoagulant activity. Samples were also assayed for specific clotting factors, including Factor XI (FXI), activated FXI (FXIa), and Factor XII (FXII). RESULTS: The purification process effectively removes any detectable procoagulant activity from the product and reduces coagulation factor content (FXI and FXII) to levels just at or below the quantification limits of the respective methods. The caprylate treatment steps of the process are critical for procoagulant removal. CONCLUSIONS: These data demonstrate consistent and robust removal of coagulation factors by the GamunexÒ-C purification process and serve to identify the caprylate precipitation and caprylate incubation steps as critical for removal of procoagulant impurities. Overall, the entire purification process from Plasma Pool to Sterile Bulk reduces FXI and FXII content by at least 5000-fold relative to IgG concentration with removal to very low or non-quantifiable levels by the two caprylate steps.
35
Serum 25-Hydroxyvitamin D Is Associated with Positive Hepatitis B Vaccine Response Ju-Suk Lee, MD, PhD1, Jin-A. Jung, MD2, Bong-Seong Kim, MD3, Hyeon Jong Jong Yang, MD4, Ja Hyeong Kim, MD5; 1Sungkyunkwan university, Changwon, South Korea, 2Dong-A University Hospital, Busan, 3 Gangneung Asan Hospital, Gangneung-Si, 4Soonchunhyang University Hospital, Seoul; Pediatric Allergy Respiratory Center, Department of Pediatrics, Soonchunhyang University Hospital, Seoul, South Korea, 5 Ulsan University Hospital, Ulsan. RATIONALE: Vitamin D deficiency or insufficiency is a relatively common and significant public health problem. Previous lots of papers have shown the relationship between vitamin D and immune functions and .Recent paper showed vitamin D deficiency was associated with poor response to active hepatitis B immunization in patients with chronic kidney disease. According to previous report, we want to examine the associations between vitamin D levels and response to hepatitis B immunization in healthy adults in large sample size. METHODS: We recruited 5025 male adult persons in one ship building company. We assessed the relationships between positive hepatitis B vaccine response and age, BMI(body mass index), hypertension, smoke, diabetes mellitus. RESULTS: We found positive relations hepatitis B vaccine response and age (P<0.01), BMI(P 50.01), smoke (P<0.01), diabetes mellitus (p<0.01). Hypertension was not associated with hapatitis B vaccine response (p50.54). CONCLUSIONS: Vitamin D levels are associated with poor response to hepatitis B immunization.
SATURDAY
32
Cytokine Profile and Clinical Correlates in Immune Deficient (HIV-infected) Infants with Severe (hypoxic) Pneumonia Robin J. Green, MD, PhD, FAAAAI1, Refiloe Masekela, MD, Cert Pulm Paed1, Dankwart Wittenberg, MD, Cert Pulm Paed1, Alta Terblanche, Cert Gastro Paed1, Paul Rheeder, PhD2, Piet Becker, PhD3, Ronald Anderson, PhD4; 1Department of Paediatrics and Child Health, University of Pretoria, South Africa, 2Division of Clinical Epidemiology, School of Health Systems and Public Health, University of Pretoria, Pretoria, South Africa, 3 Medical Research Council, South Africa, 4Medical Research Council Unit for Inflammation and Immunity, Department of Immunology, University of Pretoria and Tshwane Academic Division of National Health Laboratory Service, South Africa. RATIONALE: Severe pneumonia in infants who are HIV-infected is a common problem in many parts of the developing world, especially subSaharan Africa. What has been missing from previous studies of severe pneumonia in HIV-infected infants, however, is a description of the host inflammatory response and cytokine/chemokine profile that accompanies this disease. To describe the cytokine profiles associated with severe hypoxic pneumonia in HIV-infected infants. METHODS: In a cohort of HIV-infected children diagnosed clinically with severe hypoxic pneumonia, paired serum and sputum cytokines were tested. A control group of HIV-infected children with bronchiectasis contributed matching controls. RESULTS: A total of 100 infants (mean age 2.8 months) with a clinical diagnosis of severe hypoxic pneumonia were included in this study. IP-10 was markedly elevated in both sputum (mean 560.77pg/ml) and serum (mean 9091.14pg/ml), while IP-10 was elevated in serum (mean 39.55 pg/ml), with both these cytokines being significantly higher than in stable children with HIV-related bronchiectasis. CONCLUSIONS: This study of HIV-infected infants with severe hypoxic pneumonia suggests that IL-10 and IP-10 are associated with more severe lung disease. However, further investigation of this association is required.
33
Atheroprotection Conferred by Immunization with 15-Mer Peptide Fragments From ApoB100 Kevin Tse, MD1, Harley Tse, PhD2, Michael Shaw2, Alessandro Sette, Biol. Sci.3, Klaus Ley, MD3; 1UCSD, 2Wayne State University, 3 La Jolla Institute for Allergy and Immunology, La Jolla, CA. RATIONALE: Native LDL vaccinations of atherosclerotic-prone mice (ApoE-/-) have yielded 50% athero-reduction when administered repeatedly at regular intervals over several months. Akin to allergy immunotherapy, the presumed mechanism of this atheroprotection is thought to be due to induction of tolerance to the suspected autoantigen, oxidized LDL. Here, we report the discovery of a novel peptide found in murine ApoB100 (the lipoprotein portion of LDL) and demonstrate its ability to confer atheroprotection. METHODS: 15-mer peptide sequences spanning ApoB100 were synthesized and screened for their ability to bind tightly to MHC class II and cause T-cell proliferation after primary immunization. The candidate peptide (CP3) was used to vaccinate 5 female ApoE-/- using 50 micrograms of CP3/CFA subcutaneously in the inguinal area at 8 wks of age. Western diet was started at 10 wks of age. Repeated boosters with 25 micrograms of CP3/IFA were administered intraperitoneally at age 12, 16, 20 and 22 weeks. Mice were sacrificed at age 23 weeks and organs were harvested for analysis. PBS and irrelevant peptide were used as controls. RESULTS: CP3 treated mice showed >50% reduction in lesion size by aortic pinning when compared to PBS (p50.003) and irrelevant peptide controls (p50.004). No differences were observed in the percentage of IFNg, IL-2, IL-10, IL-17A, or FoxP3 positive cells in spleen or lymph nodes by intracellular FACS staining. CONCLUSIONS: CP3 is a novel peptide with T-cell specificity which affords atheroprotection to a similar degree as native LDL when used in a vaccination scheme.
J ALLERGY CLIN IMMUNOL FEBRUARY 2013
34
Removal of Coagulation Factors by the GamunexÒ-C Purification Process Melanie Williams, PhD, Jeff Willey, Katherine Tull, Glenn Stevenson, Mona Parks, LaToya McElrath, Luda Golovko, Catherine Russ, John Griffin, Pete Vandeberg; Grifols, Inc., Raleigh, NC. RATIONALE: GamunexÒ-C is a highly purified human plasma-derived intravenous immunoglobulin (IVIG). Coagulation factors are natural components of the starting plasma used for IVIG manufacture and are removed during purification. Residual coagulation factor activity has been associated with an increased incidence of thromboembolic adverse events with a specific immunoglobulin product. The purpose of this study was to characterize the removal of coagulation factors during the manufacture of GamunexÒ-C. METHODS: Relevant fractionation and purification process intermediates from 30 GamunexÒ-C batches were collected and analyzed for specific coagulation factors and procoagulant activity. The thrombin generation test (TGT) and the non-activated partial thromboplastin time (naPTT) are global coagulation methods that detect procoagulant activity. Samples were also assayed for specific clotting factors, including Factor XI (FXI), activated FXI (FXIa), and Factor XII (FXII). RESULTS: The purification process effectively removes any detectable procoagulant activity from the product and reduces coagulation factor content (FXI and FXII) to levels just at or below the quantification limits of the respective methods. The caprylate treatment steps of the process are critical for procoagulant removal. CONCLUSIONS: These data demonstrate consistent and robust removal of coagulation factors by the GamunexÒ-C purification process and serve to identify the caprylate precipitation and caprylate incubation steps as critical for removal of procoagulant impurities. Overall, the entire purification process from Plasma Pool to Sterile Bulk reduces FXI and FXII content by at least 5000-fold relative to IgG concentration with removal to very low or non-quantifiable levels by the two caprylate steps.
35
Serum 25-Hydroxyvitamin D Is Associated with Positive Hepatitis B Vaccine Response Ju-Suk Lee, MD, PhD1, Jin-A. Jung, MD2, Bong-Seong Kim, MD3, Hyeon Jong Jong Yang, MD4, Ja Hyeong Kim, MD5; 1Sungkyunkwan university, Changwon, South Korea, 2Dong-A University Hospital, Busan, 3 Gangneung Asan Hospital, Gangneung-Si, 4Soonchunhyang University Hospital, Seoul; Pediatric Allergy Respiratory Center, Department of Pediatrics, Soonchunhyang University Hospital, Seoul, South Korea, 5 Ulsan University Hospital, Ulsan. RATIONALE: Vitamin D deficiency or insufficiency is a relatively common and significant public health problem. Previous lots of papers have shown the relationship between vitamin D and immune functions and .Recent paper showed vitamin D deficiency was associated with poor response to active hepatitis B immunization in patients with chronic kidney disease. According to previous report, we want to examine the associations between vitamin D levels and response to hepatitis B immunization in healthy adults in large sample size. METHODS: We recruited 5025 male adult persons in one ship building company. We assessed the relationships between positive hepatitis B vaccine response and age, BMI(body mass index), hypertension, smoke, diabetes mellitus. RESULTS: We found positive relations hepatitis B vaccine response and age (P<0.01), BMI(P 50.01), smoke (P<0.01), diabetes mellitus (p<0.01). Hypertension was not associated with hapatitis B vaccine response (p50.54). CONCLUSIONS: Vitamin D levels are associated with poor response to hepatitis B immunization.