Tanto a luteína quanto o ácido linoleico conjugado (CLA) são nutrientes que já demonstraram efeitos benéficos na modulação do sistema imune em diferentes modelos experimentais, porém o uso de ambos em conjunto ainda não foi explorado. O CLA é incorporado nas membranas celulares e hipotetiza-se que a luteína, com seu potencial antioxidante, possa proteger a estrutura do CLA em situações de estresse oxidativo. Outra razão para se pensar na interação entre estes nutrientes é que já foram observados de forma individual, efeitos modulatórios do CLA e da luteína sobre receptores celulares que atuam em conjunto e possuem papel chave na regulação da resposta imune (receptores PPAR e RXR). Objetivou-se com o presente estudo avaliar o efeito da suplementação dietética de CLA e luteína no desempenho e na resposta imune de frangos de corte de 1 a 22 dias de idade. Foram testados 3 níveis de inclusão de CLA (0, 1 e 2%) em conjunto com luteína (0 e 50 mg/kg) na presença ou ausência de desafio imunológico com LPS. O CLA e a luteína apresentaram efeitos imunomodulatórios positivos, porém, não foi observada interação entre ambos os nutrientes para as avaliações relacionadas ao sistema imune. O CLA, adicionado em 2% na dieta, elevou a produção de IgY em resposta ao estímulo com albumina de soro bovino (BSA) e também foi capaz de aumentar a expressão de RXRα no fígado. A luteína diminuiu o óxido nítrico plasmático e também a expressão de TLR-4 no baço e de IL-1β e IL-12 no fígado apesar de ter aumentado a expressão de TLR-4 hepática. O desafio com LPS estimulou a resposta inflamatória aguda, evidenciado pela queda no ganho de peso, pelo aumento da relação fígado:peso corporal, pelo aumento na expressão de IL-1β e IL-12 hepáticos e diminuição na expressão de PPARα no duodeno e fígado e de PPARγ e RXRα no baço. Entretanto, nem a luteína e nem o CLA foram capazes de reverter ou atenuar os efeitos causados pelo desafio com LPS. Para desempenho, uma forte interação entre CLA e luteína foi observada, de forma que a suplementação com 1 ou 2% de CLA pioraram o peso corporal, o ganho de peso e a eficiência alimentar de 1 a 20 dias de idade, mas estes efeitos foram revertidos quando a luteína foi incluída na dieta com 1% de CLA. Concluiu-se que o CLA teve efeito benéfico sobre a resposta imune humoral, na dependência da sua dose. A luteína se mostrou como nutriente anti- inflamatório e também capaz de reverter o efeito negativo da inclusão dietética de 1% de CLA sobre o desempenho de frangos de corte. / Both lutein and conjugated linoleic acid (CLA) have beneficial effects on the modulation of the immune system. However, the simultaneous supplementation of both lutein and CLA has not yet been examined. CLA is incorporated into cell membranes, and it is hypothesized that lutein, as an antioxidant, protects CLA structure during oxidative stress. Additionally, an examination of the interaction of lutein and CLA is vital because individual effects of CLA and lutein on nuclear receptors that work together and have important roles in the immune response have been observed (PPAR and RXR receptors). The objective of this study was to evaluate the effects of dietary CLA and lutein supplementation on the performance and immune response of 1- to 22-d-old broiler chickens. Three CLA inclusion levels (0, 1 and 2%) and two lutein levels (0 and 50 mg/kg) were tested in the presence and absence of LPS immune challenge. Lutein and CLA showed positive immunomodulatory effects, but no interaction between these nutrients on the immune system was observed. A 2% CLA supplementation increased plasmatic IgY anti-BSA production and hepatic RXRα expression. Lutein decreased plasmatic nitric oxide and TLR-4 in the spleen and IL-1β and IL-12 in the liver in addition to increasing hepatic TLR-4. LPS challenge effectively promoted an acute inflammatory response, as illustrated by decreased body weight gain, increased liver:body weight ratio, increased expression of hepatic IL-1β and IL-12, decreased expression of PPARα in the duodenum and liver and decreased expression of PPARγ and RXRα in the spleen. However, neither lutein nor CLA reversed or attenuated the effects of the LPS challenge. A strong interaction between CLA and lutein was observed on performance: CLA supplementation at 1 or 2% decreased body weight, body weight gain and feed efficiency from 1 to 20 d old. However, these negative effects were reversed when lutein was included in the 1% CLA diet. In conclusion, CLA improved the humoral immune response (depending on CLA dose). Lutein was anti-inflammatory and could reverse the negative effects of dietary 1% CLA supplementation on broiler chicken performance.

Tanto a luteína quanto o ácido linoleico conjugado (CLA) são nutrientes que já demonstraram efeitos benéficos na modulação do sistema imune em diferentes modelos experimentais, porém o uso de ambos em conjunto ainda não foi explorado. O CLA é incorporado nas membranas celulares e hipotetiza-se que a luteína, com seu potencial antioxidante, possa proteger a estrutura do CLA em situações de estresse oxidativo. Outra razão para se pensar na interação entre estes nutrientes é que já foram observados de forma individual, efeitos modulatórios do CLA e da luteína sobre receptores celulares que atuam em conjunto e possuem papel chave na regulação da resposta imune (receptores PPAR e RXR). Objetivou-se com o presente estudo avaliar o efeito da suplementação dietética de CLA e luteína no desempenho e na resposta imune de frangos de corte de 1 a 22 dias de idade. Foram testados 3 níveis de inclusão de CLA (0, 1 e 2%) em conjunto com luteína (0 e 50 mg/kg) na presença ou ausência de desafio imunológico com LPS. O CLA e a luteína apresentaram efeitos imunomodulatórios positivos, porém, não foi observada interação entre ambos os nutrientes para as avaliações relacionadas ao sistema imune. O CLA, adicionado em 2% na dieta, elevou a produção de IgY em resposta ao estímulo com albumina de soro bovino (BSA) e também foi capaz de aumentar a expressão de RXRα no fígado. A luteína diminuiu o óxido nítrico plasmático e também a expressão de TLR-4 no baço e de IL-1β e IL-12 no fígado apesar de ter aumentado a expressão de TLR-4 hepática. O desafio com LPS estimulou a resposta inflamatória aguda, evidenciado pela queda no ganho de peso, pelo aumento da relação fígado:peso corporal, pelo aumento na expressão de IL-1β e IL-12 hepáticos e diminuição na expressão de PPARα no duodeno e fígado e de PPARγ e RXRα no baço. Entretanto, nem a luteína e nem o CLA foram capazes de reverter ou atenuar os efeitos causados pelo desafio com LPS. Para desempenho, uma forte interação entre CLA e luteína foi observada, de forma que a suplementação com 1 ou 2% de CLA pioraram o peso corporal, o ganho de peso e a eficiência alimentar de 1 a 20 dias de idade, mas estes efeitos foram revertidos quando a luteína foi incluída na dieta com 1% de CLA. Concluiu-se que o CLA teve efeito benéfico sobre a resposta imune humoral, na dependência da sua dose. A luteína se mostrou como nutriente anti- inflamatório e também capaz de reverter o efeito negativo da inclusão dietética de 1% de CLA sobre o desempenho de frangos de corte. / Both lutein and conjugated linoleic acid (CLA) have beneficial effects on the modulation of the immune system. However, the simultaneous supplementation of both lutein and CLA has not yet been examined. CLA is incorporated into cell membranes, and it is hypothesized that lutein, as an antioxidant, protects CLA structure during oxidative stress. Additionally, an examination of the interaction of lutein and CLA is vital because individual effects of CLA and lutein on nuclear receptors that work together and have important roles in the immune response have been observed (PPAR and RXR receptors). The objective of this study was to evaluate the effects of dietary CLA and lutein supplementation on the performance and immune response of 1- to 22-d-old broiler chickens. Three CLA inclusion levels (0, 1 and 2%) and two lutein levels (0 and 50 mg/kg) were tested in the presence and absence of LPS immune challenge. Lutein and CLA showed positive immunomodulatory effects, but no interaction between these nutrients on the immune system was observed. A 2% CLA supplementation increased plasmatic IgY anti-BSA production and hepatic RXRα expression. Lutein decreased plasmatic nitric oxide and TLR-4 in the spleen and IL-1β and IL-12 in the liver in addition to increasing hepatic TLR-4. LPS challenge effectively promoted an acute inflammatory response, as illustrated by decreased body weight gain, increased liver:body weight ratio, increased expression of hepatic IL-1β and IL-12, decreased expression of PPARα in the duodenum and liver and decreased expression of PPARγ and RXRα in the spleen. However, neither lutein nor CLA reversed or attenuated the effects of the LPS challenge. A strong interaction between CLA and lutein was observed on performance: CLA supplementation at 1 or 2% decreased body weight, body weight gain and feed efficiency from 1 to 20 d old. However, these negative effects were reversed when lutein was included in the 1% CLA diet. In conclusion, CLA improved the humoral immune response (depending on CLA dose). Lutein was anti-inflammatory and could reverse the negative effects of dietary 1% CLA supplementation on broiler chicken performance.

As respostas inflamatórias no cérebro são mediadas principalmente pela microglia, mas evidências crescentes sugerem uma importância crucial dos astrócitos. A S100B, uma proteína ligante de cálcio e secretada por astrócitos, tem propriedades neurotróficas e de citocina inflamatória. No entanto, não se sabe se sinais primários que ocorrem durante a indução de uma resposta inflamatória como, por exemplo, lipopolissacarídeo (LPS) modulam diretamente a S100B. A neuroinflamação está implicada na patogênese ou na progressão de uma variedade de distúrbios neurodegenerativos e muitos estudos procuram uma conexão entre S100B e doenças degenerativas, incluindo a doença de Alzheimer e esquizofrenia. O uso terapêutico de fármacos anti-inflamatórios não-esteroidais (AINEs) para estas doenças tem aumentado. No entanto, existem poucos estudos sobre o efeito desses fármacos em relação à proteína S100B. Neste trabalho, nós avaliamos se os níveis de S100B no líquido cefalorraquidiano (LCR) e soro de ratos Wistar são afetados por injeção de LPS administrado por via intraperitoneal (IP) ou intracerebroventricular (ICV), bem como se as culturas primárias de astrócitos respondem diretamente ao LPS. Além disso, nós avaliamos o conteúdo e a secreção de S100B medido por ELISA (bem como o conteúdo de GFAP e secreção de TNF-α) em culturas primárias de astrócitos expostos a dexametasona e quatro classes químicas diferentes de AINEs (ácido acetilsalicílico, ibuprofeno, diclofenaco e nimesulida) durante 24 h. Os nossos dados sugerem que a secreção de S100B no tecido cerebral é estimulada rapidamente e persistentemente (durante pelo menos 24 h) por administração ICV de LPS. Este aumento da S100B no LCR foi transitório quando o LPS foi administrado IP. Em contraste com estes resultados de S100B, observou-se um aumento nos níveis de TNF-α no soro, mas não no LCR, após a administração IP de LPS. Em astrócitos isolados e em fatias de hipocampo frescas, observou-se uma estimulação direta da secreção de S100B por LPS numa concentração de 10 ug/ml. Um envolvimento de TLR4 foi confirmado pelo uso de antagonistas específicos deste receptor. No entanto, baixas concentrações de LPS em culturas de astrócitos foram capazes de induzir uma diminuição na secreção de S100B após 24 h, sem alteração significativa no conteúdo intracelular de S100B. Além disso, após 24 horas de exposição ao LPS, observou-se um decréscimo na glutationa e um aumento na proteína ácida fibrilar glial. Também foi observado que os AINEs apresentam diferentes efeitos sobre parâmetros gliais. O ácido acetilsalicílico e o diclofenaco foram capazes de aumentar a GFAP, enquanto que a nimesulida, um inibidor seletivo de COX-2, e a dexametasona diminuiram a secreção de S100B. No entanto, todos os AINEs reduziram os níveis de PGE2. Juntos, esses dados contribuem para a compreensão dos efeitos de LPS em astrócitos, especialmente sobre a secreção de S100B, e nos ajuda a interpretar mudanças nesta proteína no LCR e soro em doenças neuroinflamatórias. Além disso, tecidos periféricos que expressam S100B talvez devam ser regulados diferentemente, uma vez que a administração IP de LPS não foi capaz de aumentar os níveis séricos de S100B. Em relação aos AINEs, a PGE2 parece estar envolvida no mecanismo de secreção de S100B, mas vias adicionais, não claras neste momento, necessitam de uma maior caracterização. O papel inflamatório de S100B em doenças degenerativas, onde também são observados níveis elevados da COX-2 e PGE2, poderia ser atenuado por inibidores de COX-2. / Inflammatory responses in brain are primarily mediated by microglia, but growing evidence suggests a crucial importance of astrocytes. S100B, a calciumbinding protein secreted by astrocytes, may act as a neurotrophic or an inflammatory cytokine. However, it is not known whether primary signals occurring during induction of an inflammatory response (e.g. lipopolysaccharide, LPS) directly modulate S100B. Neuroinflammation has been implicated in the pathogenesis or progression of a variety of neurodegenerative disorders and several studies have looked for a connection of S100B, and degenerative diseases including Alzheimer’s disease and schizophrenia. The therapeutic use of non-steroid anti-inflammatory drugs (NSAID) to these diseases has growth up. However, there are few reports about the effect of these drugs on S100B. In this work, we evaluated whether S100B levels in cerebrospinal fluid (CSF) and serum of Wistar rats are affected by LPS administered by intraperitoneal (IP) or intracerebroventricular (ICV) injection, as well as whether primary astrocyte cultures respond directly to lipopolysaccharide. Moreover we evaluated S100B content and secretion measured by ELISA (as well as GFAP content and TNF-α secretion) in primary astrocyte cultures exposed to dexamethasone and 4 different chemical classes of NSAID (acetyl salicylic acid, ibuprofen, diclofenac and nimesulide) for 24 h. Our data suggest that S100B secretion in brain tissue is stimulated rapidly and persistently (for at least 24 h) by ICV LPS administration. This increase in CSF S100B was transient when LPS was IP administered. In contrast to these S100B results, we observed an increase in in TNFα levels in serum, but not in CSF, after IP administration of LPS. In isolated astrocytes and in acute hippocampal slices, we observed a direct stimulation of S100B secretion by LPS at a concentration of 10 μg/mL. An involvement of TLR4 was confirmed by use of specific inhibitors. However, lower levels of LPS in astrocyte cultures were able to induce a decrease in S100B secretion after 24 h, without significant change in intracellular content of S100B. In addition, after 24 h exposure to LPS, we observed a decrease in astrocytic glutathione and an increase in astrocytic glial fibrillary acidic protein. We also observe that NSAIDs have distinct effects on glial parameters. ASA and diclofenac are able to increase GFAP, while nimesulide, a selective COX-2 inhibitor, and dexamethasone were able to decrease S100B secretion. However, all anti-inflammatories were able to reduce levels of PGE2. Together, these data contribute to the understanding of the effects of LPS on astrocytes, particularly on S100B secretion, and help us to interpret cerebrospinal fluid and serum changes for this protein in neuroinflammatory diseases. Moreover, non-brain S100B-expressing tissues may be differentially regulated, since LPS administration did not lead to increased serum levels of S100. With respect to NSAIDs, PGE2 is possibly involved in the mechanism of S100B secretion but additional pathways, unclear at this moment, demand further characterization. The inflammatory role of S100B in degenerative diseases, where also is observed elevated levels of COX-2 and PGE2, could be attenuated by COX-2 inhibitors in which elevated levels of COX-2.

IL-12, an immunoregulatory cytokine, plays a key role in the development of cell-mediated immune responses. However, very little is known about the regulation and induction of the other members of this family, particularly IL-23 and IL-27. The regulation of these cytokines was studied in the human primary monocytes and monocyte-derived macrophages (MDMs) as they play a key role in innate and adaptive immune responses. THP-1 promonocytic cells were employed as a model system to confirm the results obtained with monocytes and MDMs. Two stimuli IFN-γ and LPS were used as both are strong inducers of IL-12 family cytokines. My results show that IFN-γ induced the production of IL-12/23p40 and IL-23p19 mRNA as well as IL-12p40 and IL-23 proteins in primary human monocytes isolated by positive selection. IFN-γ-induced IL-23 and IL-12/23p40 expression was positively regulated by the p38 mitogen-activated protein kinases (MAPK), independent of the Janus kinase (Jak)/signal transducers and activators of transcription (STAT) signaling. In contrast, IL-12 and IL-23 were negatively regulated by the Jak/STAT, phosphoinositide-3 kinase (PI3K) and the c-Jun-N-terminal kinase (JNK) MAPKs in IFN-γ-stimulated monocytes. LPS significantly stimulated IL-23p19 and IL-12/23p40 mRNA expression as well as IL-12/23p40 and IL-23 protein production in THP-1 cells, while IFN-γ stimulation alone did not affect IL-23 mRNA or protein levels. THP-1 cells were pre-treated with ERK, JNK or p38 MAPK inhibitors and then stimulated with LPS. LPS-induced IL-12p40 and IL-23 proteins were positively regulated by the p38 and JNK MAPKs and PI3K, whereas LPS-induced IL-23p19 mRNA expression was negatively regulated by these kinases. These results were confirmed using siRNA in LPS-stimulated THP-1 cells. My results also show that IFN-γ/LPS-induced IL-23 expression is not regulated through MAPK or PI3K signaling pathways in human MDMs. My results also show for the first time that IFN-γ alone without any second stimulus induced IL-27p28 gene expression and IL-27 protein production in human monocytic cells. I investigated the signalling pathways governing the regulation of IL-27 protein and its subunit IL-27p28 following stimulation with IFN-γ in primary human monocytic cells. IFN-γ-mediated IL-27 protein, but not IL-27p28 gene expression was positively regulated by JNK MAPK and PI3K, independent of JAK/STAT signaling in primary human monocytes. I also investigated the signalling pathways governing the regulation of IL-27 and its α subunit, IL-27p28 following stimulation with IFN-γ alone or IFN-γ-primed LPS-stimulated macrophages (IFN-γ/LPS) and THP-1 cells. A differential regulation of IL-27p28 and IL-27 in response to stimulation by either IFN-γ or IFN-γ/LPS was observed. IFN-γ- and IFN-γ/LPS induced IL-27 expression was positively regulated by the JNK, p38 MAPK and PI3K, independent of Jak/STAT signaling in human MDMs and THP-1 cells. Taken together, my results show that IL-23 induction is differentially regulated by different pathways in response to different stimuli, whereas IL-27 expression is regulated by JNK, p38 MAPK and PI3K regardless in the stimulus in human myeloid cells. These results may provide additional strategies aimed at targeting disease, autoimmune disorders and cancer.

Inflammation plays a crucial role in protecting the host from infection and tissue injury. However, uncontrolled inflammation contributes to the pathogenesis of major auto-inflammatory diseases. Interleukin-1 (IL-1), a pleiotropic pro-inflammatory cytokine, is a pivotal mediator of many of these diseases. The best characterised IL-1 family members, IL-1α and IL-1β, are produced as precursor forms of 31 kDa in size. Both precursors are cleaved and secreted, activating transmembrane IL-1 receptors on IL-1-responsive cells. Many studies that focused on IL-1α have shown that the precursor and processed mature Ct peptide, as well as its N terminus (Nt) form, can elicit a signal. However, with IL-1β, only the processed mature Ct form is known to elicit an inflammatory response and no immunological activity has been attributed to Nt fragments of pro-IL-1β. Therefore, the first objective of this study was to produce recombinant human Nt-IL-1β fragments in bacterial and mammalian expression system to investigate their possible immunomodulatory functions. Recombinant His-tagged N-terminus fragments (10 and 14 kDa) of pro-IL-1β were cloned into the bacterial expression vector pET-22(+) and expressed in E. coli BL21(DE3) followed by purification using three consecutive columns (IMAC, SEC and AEC). Purification analysis of eluted proteins from columns indicated that the recombinant proteins were always co-purified with some other bacterial proteins. The Nt fragments of pro-IL-1β were cloned into the mammalian expression plasmid, pcDNA3.1(+). Expression of these proteins was monitored by transfection of two mammalian cell lines: Human Embryonic Kidney (HEK) 293 cells and monkey kidney cells (COS-7). No protein expression was observed with either construct. These limitations urged us to investigate the expression and degradation of endogenous IL-1 in vitro. Previous studies have shown that the transcription of cytokine genes in response to lipopolysaccharide (LPS) is usually rapid and begins to decline within a few hours after stimulation. The proteasome is the major cellular proteolytic apparatus and controls the turn-over of cellular proteins. We investigated the intracellular stability of IL-1α and IL-1β in LPS-stimulated mouse J774 macrophages and primary mouse bone marrow derived macrophages (BMDMs). Exposure of LPS-stimulated J774 and BMDMs to three different classes of proteasome inhibitors (peptide alhedyde (ALLN), peptide boronate (MG262) and non-peptide inhibitor (β-lactone)) prevented the degradation of intracellular IL-1α and IL-1β in a concentration and time dependent manner. Furthermore, the release of IL-1 into the culture media was not affected by any of these inhibitors in LPS-stimulated J774 cells. However, in LPS-stimulated BMDMs, β-lactone increased the release of both IL-1α and IL-1β and ALLN only increased IL-1α release into culture supernatant compared to control. MG262 had no effect on the release of either. These data suggest that the proteasome plays an important role in controlling the amount of IL-1α and IL-1β by restricting the intracellular levels of these cytokines in activated monocytes and macrophages. Therefore, this study provides evidence in support of the hypothesis that the proteasome is involved in the degradation of IL-1α and IL-1β and may offer a potential therapeutic target in inflammatory diseases.

Confinement of the obligate intracellular bacterium Chlamydia trachomatis to a membrane-bound vacuole, termed an inclusion, within infected epithelial cells neither prevents secretion of chlamydial antigens into the host cytosol nor protects chlamydiae from innate immune detection. However, the details leading to chlamydial antigen presentation are not clear. By immunoelectron microscopy of infected endometrial epithelial cells and in isolated cell secretory compartments, chlamydial major outer membrane protein (MOMP), lipopolysaccharide (LPS) and the inclusion membrane protein A (IncA) were localized to the endoplasmic reticulum (ER) and co-localized with multiple ER markers, but not with markers of the endosomes, lysosomes, Golgi nor mitochondria. Chlamydial LPS was also co-localized with CD1d in the ER. Since the chlamydial antigens, contained in everted inclusion membrane vesicles, were found within the host cell ER, these data raise additional implications for antigen processing by infected uterine epithelial cells for classical and non-classical T cell antigen presentation.

Lipopolysaccharide or LPS is localized to the outer leaflet of the outer membrane and serves as the major surface component of the bacterial cell envelope. This remarkable glycolipid is essential for virtually all Gram-negative organisms and represents one of the conserved microbial structures responsible for activation of the innate immune system. For these reasons, the structure, function, and biosynthesis of LPS has been an area of intense research. The LPS of a number of bacteria is composed of three distinct regions - lipid A, a short core oligosaccharide, and the O-antigen polysaccharide. The lipid A domain, also known as endotoxin, anchors the molecule in the outer membrane and is the bioactive component recognized by TLR4 during human infection. Overall, the biochemical synthesis of lipid A is a highly conserved process; however, investigation of the lipid A structures of various organisms shows an impressive amount of diversity. These differences can be attributed to the action of latent enzymes that modify the canonical lipid A molecule. Variation of the lipid A domain of LPS serves as one strategy utilized by Gram-negative bacteria to promote survival by providing resistance to components of the innate immune system and helping to evade recognition by TLR4. This review summarizes the biochemical machinery required for the production of diverse lipid A structures of human pathogens and how structural modification of endotoxin impacts pathogenesis.

No description available.

For a long time, bilirubin (BR) has been considered a waste molecule with potential toxic effects especially on the central nervous system. Later, it was found that BR exhibited cytoprotective effects and mildly elevated BR levels showed antioxidant, anti-inflammatory and immunomodulatory properties, however, exact mechanisms of the anti-inflammatory actions of BR have not been fully understood yet. The main aim of this study was to assess the protective effects of BR using experimental in vivo and in vitro models in relation to inflammation and oxidative stress. Partial goal was to establish validated analytical method for determination of BR and lumirubin. Gunn and heterozygous rats were treated with lipopolysaccharide (LPS, 6 mg/kg, IP) or vehicle (saline). After 12 hours, blood and organs were collected for analyses of inflammatory and hepatic injury markers. Primary rat hepatocytes were treated with BR and TNF-α, HepG2 and SH-SY5Y cell lines were treated with BR and chenodeoxycholic acid. LPS-treated Gunn rats had a significantly decreased inflammatory response and hepatic injury compared to LPS- treated normobilirubinemic controls. We found different profile of leukocytes subsets and decreased systemic mRNA expressions and concentrations of IL-6, TNF-α, IL-1β and IL-10 in Gunn rats. Hepatic mRNA...

Peripubertal immune stress alters adult responsiveness to estradiol (E2) and progesterone (P). When female mice are injected with the bacterial endotoxin lipopolysaccharide (LPS) at six weeks of age, or during pubertal development, they display a decrease in response to ovarian hormones. In contrast, females ovariectomized prior to peripubertal immune stress display typical levels of sexual behavior following sequential injections of E2 and P in adulthood. Additionally, intact females exposed to peripubertal immune stress display a decrease in estrogen receptor alpha (ER-α)-immunoreactive (ir) cells in the medial preoptic area (MPOA) and ventromedial nucleus of the hypothalamus (VMH) in adulthood. However, ER-α has not been studied in mice that have been ovariectomized prior to receiving LPS. The objective of the present study is two-fold: to replicate the finding that ovariectomy prior to pubertal development prevents the deleterious effects of LPS administration, and to examine the status of ER-α in areas of the brain important to sex behavior. We predicted that mice ovariectomized after LPS injection would display fewer ER-α-ir cells and a decreased responsiveness to ovarian hormones than saline controls and those mice ovariectomized prior to LPS injection. To test this, female mice were ovariectomized or sham-operated prior to LPS treatment. Then, at six weeks of age, all mice were injected with saline or LPS. Following that, sham-operated mice were ovariectomized and ovariectomized mice were sham-operated. Mice were primed weekly with E2 and P, and sex behavior testing occurred once a week for 5 weeks. After the final behavior test, all mice were euthanized, their brains removed, and stained for ER-α via immunocytochemistry. Results revealed a large variability in hormone responsiveness. However, animals that received peripubertal LPS, but still had their ovaries, had significantly lower sexual receptivity when compared to animals that were ovariectomized prior to the pubertal period and given LPS. Further, there were no differences between groups in ER-α-ir numbers. External environmental stressors, such as animal housing and vibrations and noise from nearby construction, may have caused some of the results found here, which are inconsistent with previous findings.