The drug-drug interaction involving zidovudine and sulfamethoxazole-trimethoprim was investigated using an in vitro culture system, an in vivo mouse model, and a clinical trial in HIV-infected patients. We hypothesized that combination exposure causes immune cell populations in the bone marrow to undergo apoptotic cell death, and that the toxicity would affect the host response to an infectious stimulus. Mice were dosed with zidovudine, sulfamethoxazole-trimethoprim, the combination of both drugs, or vehicle only control via oral gavage. Focusing on B-lineage cells in the bone marrow, we determined that cells of the rapidly cycling, early pre-B cell subset are targeted, as well as pro-B cells earlier in development. This toxicity was found to be cell cycle dependent, with an increase in percentage of cells in the S/G2/M phases of the cycle. In vitro experiments using the drugs in a bone marrow culture system demonstrated that the effect of cytotoxicity with combination exposure is synergistic and concentration-dependent. The mechanism of apoptosis that is induced appears to be caspase-independent. To measure host response in mice, animals treated with zidovudine plus sulfamethoxazole-trimethoprim were infected with Pneumocystis murina pneumonia, and the group that received the combination of agents had a blunted antigen-specific IgG response, possibly due to a decreased number of B cells and activated B cells in the draining lymph nodes of the lungs. A clinical trial was conducted in HIV-infected patients, dividing subjects into groups receiving zidovudine, sulfamethoxazole-trimethoprim, the combination of both, or neither agent. Upon vaccination with the influenza vaccine, the combination treatment group had a blunted humoral response, with reduced antigen-specific serum IgG titers as compared to the control group. We conclude that the drug-drug interaction involving zidovudine and sulfamethoxazole-trimethoprim is clinically-significant, and clinicians must consider this toxicity when treating patients with these agents concurrently.

L2pB1 cells are a subset of B-1 B lymphocytes expressing programmed death ligand 2 (PD-L2) on their surface. They constitute 30-50% of B lymphocytes in the mouse peritoneal cavity and contribute to the production of natural IgM antibody. Previous studies have indicated a protective role of B-1 B cells in attenuating atherosclerosis and insulin resistance. We report that L2pB1 cells possess a unique IgM antibody specificity for phosphorylcholine (PC) and phosphatidylcholine (PtC) IgM enabling them to perform PC- and PtC-specific phagocytosis of PtC-nanoparticles. Here we demonstrate that induced depletion of L2pB1 in a transgenic mouse model with L2pB1-specific diphtheria toxin receptor (DTR) expression increases abdominal inflammation in the peritoneal cavity as well as the visceral adipose tissue in diet-induced obese (DIO) mice. L2pB1-depleted DIO mice also display increased triglycerides and blood glucose levels per gram of body weight relative to PBS-injected control DIO mice. Our results suggest that L2pB1 cells may play a role in anti-inflammatory regulation in DIO. Further investigation is required to discover how L2pB1 cells protect from obesity-induced inflammation and whether L2pB1 cells can provide cellular therapy to control chronic inflammation in obese patients. / 2018-06-16T00:00:00Z

A imunodeficiência comum variável (CVID) é caracterizada por hipogamaglobulinemia. Anteriormente identificou-se uma paciente com CVID que apresenta nível aumentado de estresse de retículo endoplasmático (ER), secundário a desregulação da via UPR. No presente trabalho, estendemos esta análise para outros pacientes e avaliamos o perfil de maturação de seus linfócitos B. Métodos: Western-blot, RT-PCR, Q-PCR, Citometria de Fluxo e cultura de células B ex vivo e imortalizadas. Resultados: A análise de 16 pacientes com CVID e 9 indivíduos saudáveis revelou três pacientes com porcentagens aumentadas de linfócitos B imaturos no sangue periférico. A análise da expressão de RNAm para BiP e XBP-1 em linfócitos B destes pacientes, após estímulo com LPS in vitro, identificou que os linfócitos B de um deles apresenta estresse de RE. Conclusão: Identificamos um subgrupo de pacientes com CVID que apresentam linfócitos B imaturos no sangue periférico. Um membro deste subgrupo apresenta estresse aumentado de ER. / Common Variable Immunodeficiency (CVID) is characterized by hypogammaglobulinemia. Previously a CVID patient was identified with increased levels of Endoplasmic Reticulum (ER) stress due to dysregulation of the UPR. In the present study these analyses were performed in other patients and healthy donors. Maturation markers of B lymphocytes were also characterized in these individuals. Methods: Western-blot, RT-PCR, Q-PCR, Flow cytometry and culturing of ex vivo and immortalized B cells. Results: The analysis of 16 CVID patients and 9 healthy donors revealed three patients that present higher percentage of immature B cells in peripheral blood. Analysis of expression of BiP and XBP1 induced by LPS treatment of B lymphocytes from these patients revealed that one patient present increased levels of ER stress.

Diss. (sammanfattning) Göteborg : Univ. , 2007. / Härtill 3 uppsatser.

Diss. (sammanfattning) Göteborg : Univ., 2008. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 5 uppsatser.

Diss. (sammanfattning) Uppsala : Univ., 2001. / Härtill 6 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst. / Härtill 5 uppsatser.

Thesis (Ph. D.)--Cornell University, 1989. / Vita. Includes bibliographical references.

Thesis (Ph.D.)--University of Tennessee Health Science Center, 2008. / Title from title page screen (viewed on September 9, 2008). Research advisor: Marko Z. Radic, Ph.D. Document formatted into pages (xi,111 p. : ill.). Vita. Abstract. Includes bibliographical references (p. 91-111).