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Effects of sleep disorders and fatigue on HIV disease progression in older children and adolescents
Abstracts S223
J ALLERGY CLIN IMMUNOL VOLUME 115, NUMBER 2
New Adjuvant for Immunotherapy: Gantrez® AN Nanoparticles S. Gomez1, C. Carlos2, E. Luquin3, B. San Roman1, S. Espuelas1, M. L. Sanz3, M. Ferrer3, J. M. Irache1; 1Pharmaceutical Technology, Universidad de Navarra, Pamplona, SPAIN, 2Microbiology, Universidad de Navarra, Pamplona, SPAIN, 3Allergy and Clinical Immunology, Universidad de Navarra, Pamplona, SPAIN. RATIONALE: Gantrez® AN were examined as immunoadjuvant studying their protective effect in sensitized mice with ovalbumin. METHODS: Ovalbumin-loaded nanoparticles were prepared by a solvent displacement method. In some cases, nanoparticles were prepared in the presence of rough lipopolysacharide (R-LPS) as immunomodulator (R-LPS-NP). BALB/c mice were sensitized by treatment with 50 g ovalbumin adsorbed in alhydrogel on days 0 and 7th. On day 13th , 16th and 19th the animals received i.d. doses of different nanoparticle formulations. On day 34th, mice were challenged by i.p. administration of 1 mg ovalbumin in PBS. The intensity of the anaphylaxis was evaluated determining mortality or symptomatology, and by quantification of the histamine blood level and body temperature. RESULTS: Gantrez® AN nanoparticles displayed a size of about 200 nm and were able to carry about 30 g OVA/mg nanoparticle. The presence of R-LPS did not influence the physicochemical characteristics. In vivo, mice treated with nanoparticles containing R-LPS (R-LPS-NP) displayed the most favorable symptomatology. In fact, this group of mice showed a significantly lower capacity to rise the histamine blood level after the challenge. Similarly, the decrease on the body temperature was almost inappreciable. In addition, all the animals that treated were protected against death by anaphylactic shock. CONCLUSIONS: Gantrez® AN nanoparticles containing R-LPS protect against the anaphylactic shock in experimentally sensitized mice with ovalbumin, which encourages the application of these nanoparticles in immunotherapy. Funding: Natl Dept Science and Technology # SAF 2001-0690-C03-01
886
Effects of Oral Ingestion of Nigella sativa Seeds (Black Cumin) on Murine Immune System A. K. Y. Daoud; Internal Medicine, Jordan University Science and Technology, Irbid, JORDAN. RATIONALE: We wanted to study the effects on the murine immune system for oral ingestion of Nigella sativa seeds, which is a common herb from the family Raununuculaceae used as a food spice and thought to have many medicinal values. METHODS: 3 groups of 18 BALB/c mice were given daily either 0, 40gms or 100gms of the seeds and complete consumption was assured. The following tests were done after 2 and 3 weeks of ingestion and at 1 week after stopping the intake: - (1) White Blood Cell Count and differential,(2) NBT test on peritoneal macrophages stimulated by PMA, (3) IgM and IgG Plaque Forming Cell assays on splenocytes using sRBC as antigen and (4) Mitogenic response of splenocytes to Lipopolysaccaride (LPS) and Concanavalin A (Con A). Appropriate statistical analysis was applied. RESULTS: There was no difference between the groups for the WBC’s or differential at all times. NBT test and IgG PFC assay were significantly higher at 3 weeks and after stopping the intake with increasing doses. IgM PFC assay was significantly higher from week 2 and on with increasing doses. LPS and Con A induced proliferations were significantly higher at all times and doses. CONCLUSIONS: Nigella sativa seeds have diffuse murine immune system stimulant effects and further studies are needed on the human immune system with clarification and isolation of the active components from them.
887
A Comparison of Lymphocyte Responses in PBMC, by Human and Chimpanzees in Response to PMA/I Stimulation, or AntiCD3/Anti-CD28 Costimulation, Using Microarray Analysis J. A. DeVoti; Allergy/Immunology, North Shore-LIJ Health System, New Hyde Park, NY. RATIONALE: Although the AIDS epidemic continues to claim millions of lives annually, there is a paucity of novel therapeutic approaches being explored. The completion of the genomic sequence of the chimpanzee has provided a unique opportunity to explore the genetic basis for the resistance of our closest evolutionary relative to develop AIDS. METHODS: We used the Affymetrix microarray gene chip to explore the differences in gene expression between human and chimpanzee PBMC following in-vitro stimulation with Phorbol ester and Ionomycin or monoclonal antibodies specific for CD3 and CD28 which activate T-cells through the CD3-TCR complex and the co-stimulatory molecule CD28. RESULTS: We found a high degree of concordance between the human and chimpanzee gene expression profiles regardless of the type of stimulation used. In addition, the levels of induction for individual genes were very similar for the two species, with very little intra-, and inter-species variation. CONCLUSIONS: The remarkably similar profiles of human and chimpanzee gene expression suggest that the overall program of immunocyte gene regulation has been highly conserved between these species, despite the larger than expected differences in DNA sequences of the chimpanzee and human genomes. Thus, relatively few key differences in HIV-induced gene regulation may be responsible for the drastic differences in clinical outcome in these two genetically similar species following HIV-1 infection. Identifying novel regulatory differences in chimpanzees may provide a rationale for new therapeutic approaches in humans. Funding: NIH
888
Effects of Sleep Disorders and Fatigue on HIV Disease Progression in Older Children and Adolescents M. E. Paul1, P. Y. Brouwers1, D. G. Glaze1, J. M. Reuben2, B. Lee2, L. L. Harris1, C. A. Kozinetz1, M. W. Kline1, H. L. Schwarzwald1, S. B. Foster1, C. D. Jackson1, W. T. Shearer1; 1Pediatrics, Baylor College of Medicine, Houston, TX, 2Molecular Biology, The University of Texas, M.D. Anderson Cancer Center, Houston, TX. RATIONALE: Children with chronic illnesses have sleep disruption, daytime attention deficits, and impulsivity, and altered blood leukocyte subtypes, sleep-regulating hormones, and pro- and anti-inflammatory cytokines. Children with HIV infection exhibit similar sleep disorders and fatigue, but their effects upon HIV disease progression and immunity are unknown. METHODS: A cohort of 30 HIV+ and 30 HIV-exposed children (8-18 yr.) with detailed clinical database was secured for study. Previously validated questionnaires and test instruments (sleep habits, daytime sleepiness, neurocognition) were administered to HIV+ and control children. Blood from these children was activated with phorbol myristate acetate (PMA) or lipopolysaccharide (LPS) to assess tumor necrosis factor-alpha (TNF-a, pro-inflammatory, sleep-inducing cytokine) and interleukin-10 (IL-10, anti-inflammatory, sleep-inhibiting cytokine) expression in activated T cells to determine the relationship between sleep disturbance, fatigue, and cytokines levels. RESULTS: Psychological testing revealed that there was a positive relationship between plasma IL-10 and neurocognitive functioning in 30 HIV+ children (r = 0.40, p < 0.05). A study of a subgroup of 11 HIV+ children and 10 controls showed that TNF-a was elevated in PMA-stimulated CD3+ T cells (p = 0.049), and IL-10 was lower in LPS-stimulated CD3+ T cells (p = 0.008). CONCLUSIONS: Children with HIV infection demonstrate higher blood levels of the sleep-inducing cytokine, TNF-a; reduced blood levels of the sleep-inhibiting cytokine, IL-10; and neurocognitive deficits. These early results of this clinical study indicate that it may be possible to determine if sleep disorders and fatigue are related with disease progression in HIV+ children. Funding: NIH, NHLBI HL079533
889
TUESDAY
basophils from patients allergic to Lollium. This observation allows further research in this new way to carry the allergen. Funding: National Dept Science and Technology # SAF 2001-0690C03-01
J ALLERGY CLIN IMMUNOL VOLUME 115, NUMBER 2
New Adjuvant for Immunotherapy: Gantrez® AN Nanoparticles S. Gomez1, C. Carlos2, E. Luquin3, B. San Roman1, S. Espuelas1, M. L. Sanz3, M. Ferrer3, J. M. Irache1; 1Pharmaceutical Technology, Universidad de Navarra, Pamplona, SPAIN, 2Microbiology, Universidad de Navarra, Pamplona, SPAIN, 3Allergy and Clinical Immunology, Universidad de Navarra, Pamplona, SPAIN. RATIONALE: Gantrez® AN were examined as immunoadjuvant studying their protective effect in sensitized mice with ovalbumin. METHODS: Ovalbumin-loaded nanoparticles were prepared by a solvent displacement method. In some cases, nanoparticles were prepared in the presence of rough lipopolysacharide (R-LPS) as immunomodulator (R-LPS-NP). BALB/c mice were sensitized by treatment with 50 g ovalbumin adsorbed in alhydrogel on days 0 and 7th. On day 13th , 16th and 19th the animals received i.d. doses of different nanoparticle formulations. On day 34th, mice were challenged by i.p. administration of 1 mg ovalbumin in PBS. The intensity of the anaphylaxis was evaluated determining mortality or symptomatology, and by quantification of the histamine blood level and body temperature. RESULTS: Gantrez® AN nanoparticles displayed a size of about 200 nm and were able to carry about 30 g OVA/mg nanoparticle. The presence of R-LPS did not influence the physicochemical characteristics. In vivo, mice treated with nanoparticles containing R-LPS (R-LPS-NP) displayed the most favorable symptomatology. In fact, this group of mice showed a significantly lower capacity to rise the histamine blood level after the challenge. Similarly, the decrease on the body temperature was almost inappreciable. In addition, all the animals that treated were protected against death by anaphylactic shock. CONCLUSIONS: Gantrez® AN nanoparticles containing R-LPS protect against the anaphylactic shock in experimentally sensitized mice with ovalbumin, which encourages the application of these nanoparticles in immunotherapy. Funding: Natl Dept Science and Technology # SAF 2001-0690-C03-01
886
Effects of Oral Ingestion of Nigella sativa Seeds (Black Cumin) on Murine Immune System A. K. Y. Daoud; Internal Medicine, Jordan University Science and Technology, Irbid, JORDAN. RATIONALE: We wanted to study the effects on the murine immune system for oral ingestion of Nigella sativa seeds, which is a common herb from the family Raununuculaceae used as a food spice and thought to have many medicinal values. METHODS: 3 groups of 18 BALB/c mice were given daily either 0, 40gms or 100gms of the seeds and complete consumption was assured. The following tests were done after 2 and 3 weeks of ingestion and at 1 week after stopping the intake: - (1) White Blood Cell Count and differential,(2) NBT test on peritoneal macrophages stimulated by PMA, (3) IgM and IgG Plaque Forming Cell assays on splenocytes using sRBC as antigen and (4) Mitogenic response of splenocytes to Lipopolysaccaride (LPS) and Concanavalin A (Con A). Appropriate statistical analysis was applied. RESULTS: There was no difference between the groups for the WBC’s or differential at all times. NBT test and IgG PFC assay were significantly higher at 3 weeks and after stopping the intake with increasing doses. IgM PFC assay was significantly higher from week 2 and on with increasing doses. LPS and Con A induced proliferations were significantly higher at all times and doses. CONCLUSIONS: Nigella sativa seeds have diffuse murine immune system stimulant effects and further studies are needed on the human immune system with clarification and isolation of the active components from them.
887
A Comparison of Lymphocyte Responses in PBMC, by Human and Chimpanzees in Response to PMA/I Stimulation, or AntiCD3/Anti-CD28 Costimulation, Using Microarray Analysis J. A. DeVoti; Allergy/Immunology, North Shore-LIJ Health System, New Hyde Park, NY. RATIONALE: Although the AIDS epidemic continues to claim millions of lives annually, there is a paucity of novel therapeutic approaches being explored. The completion of the genomic sequence of the chimpanzee has provided a unique opportunity to explore the genetic basis for the resistance of our closest evolutionary relative to develop AIDS. METHODS: We used the Affymetrix microarray gene chip to explore the differences in gene expression between human and chimpanzee PBMC following in-vitro stimulation with Phorbol ester and Ionomycin or monoclonal antibodies specific for CD3 and CD28 which activate T-cells through the CD3-TCR complex and the co-stimulatory molecule CD28. RESULTS: We found a high degree of concordance between the human and chimpanzee gene expression profiles regardless of the type of stimulation used. In addition, the levels of induction for individual genes were very similar for the two species, with very little intra-, and inter-species variation. CONCLUSIONS: The remarkably similar profiles of human and chimpanzee gene expression suggest that the overall program of immunocyte gene regulation has been highly conserved between these species, despite the larger than expected differences in DNA sequences of the chimpanzee and human genomes. Thus, relatively few key differences in HIV-induced gene regulation may be responsible for the drastic differences in clinical outcome in these two genetically similar species following HIV-1 infection. Identifying novel regulatory differences in chimpanzees may provide a rationale for new therapeutic approaches in humans. Funding: NIH
888
Effects of Sleep Disorders and Fatigue on HIV Disease Progression in Older Children and Adolescents M. E. Paul1, P. Y. Brouwers1, D. G. Glaze1, J. M. Reuben2, B. Lee2, L. L. Harris1, C. A. Kozinetz1, M. W. Kline1, H. L. Schwarzwald1, S. B. Foster1, C. D. Jackson1, W. T. Shearer1; 1Pediatrics, Baylor College of Medicine, Houston, TX, 2Molecular Biology, The University of Texas, M.D. Anderson Cancer Center, Houston, TX. RATIONALE: Children with chronic illnesses have sleep disruption, daytime attention deficits, and impulsivity, and altered blood leukocyte subtypes, sleep-regulating hormones, and pro- and anti-inflammatory cytokines. Children with HIV infection exhibit similar sleep disorders and fatigue, but their effects upon HIV disease progression and immunity are unknown. METHODS: A cohort of 30 HIV+ and 30 HIV-exposed children (8-18 yr.) with detailed clinical database was secured for study. Previously validated questionnaires and test instruments (sleep habits, daytime sleepiness, neurocognition) were administered to HIV+ and control children. Blood from these children was activated with phorbol myristate acetate (PMA) or lipopolysaccharide (LPS) to assess tumor necrosis factor-alpha (TNF-a, pro-inflammatory, sleep-inducing cytokine) and interleukin-10 (IL-10, anti-inflammatory, sleep-inhibiting cytokine) expression in activated T cells to determine the relationship between sleep disturbance, fatigue, and cytokines levels. RESULTS: Psychological testing revealed that there was a positive relationship between plasma IL-10 and neurocognitive functioning in 30 HIV+ children (r = 0.40, p < 0.05). A study of a subgroup of 11 HIV+ children and 10 controls showed that TNF-a was elevated in PMA-stimulated CD3+ T cells (p = 0.049), and IL-10 was lower in LPS-stimulated CD3+ T cells (p = 0.008). CONCLUSIONS: Children with HIV infection demonstrate higher blood levels of the sleep-inducing cytokine, TNF-a; reduced blood levels of the sleep-inhibiting cytokine, IL-10; and neurocognitive deficits. These early results of this clinical study indicate that it may be possible to determine if sleep disorders and fatigue are related with disease progression in HIV+ children. Funding: NIH, NHLBI HL079533
889
TUESDAY
basophils from patients allergic to Lollium. This observation allows further research in this new way to carry the allergen. Funding: National Dept Science and Technology # SAF 2001-0690C03-01