Search

Sequential gene expression of two type 1 cytokines (interleukin 2 [IL-2] and gamma interferon), one type 2 cytokine (IL-10), two monokines (IL-6 and tumor necrosis factor alpha), and one cytokine receptor (IL-2 receptor [IL-2R]) in normal human peripheral blood mononuclear cells (PBMC) following in vitro stimulation was investigated by reverse transcription-PCR methods. Two stimuli were utilized: phytohemagglutinin (PHA), which acts on the CD2 molecule and T-cell receptors, and anti-CD3 monoclonal antibody, which acts on the CD3 molecule and on T-cell receptors. Increased expression of all studied genes occurred between 1 and 4 hours after stimulation, except for that of the gene encoding IL-10, which was delayed. Expression of all but one of the genes was transient, with a maximal mRNA accumulation at about 8 h on average. IL-2R mRNA expression was an exception, showing a prolonged increase (72 h). The general profiles of expression of the five cytokine genes were similar but not identical, suggesting some shared regulatory mechanisms. When responses to four additional stimuli (pokeweed mitogen, Candida albicans , and IL-2 at high and low doses) were compared, similar profiles of cytokine gene expression were found. Thus, the various stimuli caused induction of all cytokines with quantitative, not qualitative, differences. Altogether, the present data are useful for defining the kinetics of gene expression for key cytokines in response to standard immune-cell stimuli.

IFN-gamma mRNA levels were measured in unstimulated PBMC and purified cell subpopulations, utilizing branched DNA assays, to characterize the cell type(s) that contribute to the in vivo increase in IFN-gamma gene expression seen in HIV infection. PBMC and CD8 T cells from HIV-seropositive subjects (HIV+) showed 2.5-fold increases in mean IFN-gamma mRNA levels compared to HIV-uninfected subjects (HIV-). Within individuals, CD8 T cells showed the highest IFN-gamma expression regardless of HIV status, which suggests that HIV infection enhances the IFN-gamma gene expression in CD8 T cells rather than inducing a shift to and/or increasing expression of IFN-gamma mRNA in other cell types. HIV+ subjects with increased PBMC IFN-gamma mRNA had elevated plasma levels of HIV RNA, neopterin, and beta 2-microglobulin. No differences in IFN-gamma mRNA levels were seen among HIV+ stratified by CD4 T cell number. Increased IFN-gamma may result from or be a contributing factor to increased viral load.

OBJECTIVE: Most data on local excisions for rectal cancer are based on retrospective studies. We review the results of a prospective registry of patients eligible for local excision of rectal cancer using a transanal, transsphincteric, or transcoccygeal technique combined with multimodality therapy for lesions penetrating the muscularis propria (T2) or perirectal fat (T3). METHODS: Patients with lesions less than 4 cm in diameter and less than 10 cm from the dentate line, with no evidence of distant metastases or invasion into the perirectal fat, were eligible for local excision. Patients with invasion into the muscularis propria (T2) or greater (T3) received adjuvant chemoradiation therapy. RESULTS: Forty-eight patients have been followed prospectively. Average age is 63 years. Thirty-three patients underwent a transanal excision. Fifteen patients underwent either a transsphincteric or technique excision. There was no perioperative mortality. Pathology revealed 1 Tis, 21 T1, 21 T2, and 5 T3 cancers. Mean follow-up is 40.5 months. Cancerrelated overall mortality was 4 percent. Overall local or distant recurrence rate was 8 percent(4/48). Recurrence appeared to be related to presence of a positive margin or aggressive histology (lymphatic invasion). Local recurrences were treated with salvage therapy. CONCLUSION: Local excision can be used selectively for small rectal cancers, with minimum morbidity. Recurrence rates are low (8 percent). Patients with either a positive margin or lymphatic invasion need to be considered for further therapy, including abdominoperineal resection, even with T1 lesions. Adjuvant chemoradiation appears to be a benefit for all T2 or T3 cancers.

Social isolation is a psychosocial risk factor for age-related disease morbidity and mortality, with inflammation being a mediator in this relationship. It has also been related to a reduction in anti-inflammatory glucocorticoid-induced genes and an increased expression of pro-inflammatory molecules such as NF-κB. These previous findings also indicated activations of the JAK/STAT pathway suggesting the importance of STATs in the pro-inflammatory response. We hypothesized that socially isolated older adults would exhibit a marked difference in STAT signaling after direct cellular stimulation challenge with known pro-inflammatory cytokines. Males and females (ages 60–84), who were socially integrated ( n = 76) or socially isolated ( n = 56) in subjective feelings of social isolation, were recruited using random telephone survey of over 1600 older adults in nearby UCLA communities. Multivariate analyses examined intra-nuclear signaling of STAT1, STAT3, and STAT5 after stimulation. The results show that socially isolated older adults have higher levels of STAT5 in PBMCs, monocytes, and lymphocytes ( p p = 0.055) as compared to levels in socially integrated elderly, adjusted for age and sex. No group differences in stimulated levels of STAT3 were found. These findings support the hypothesis that social isolation primes immune cells to have an enhanced inflammatory response through STAT signaling. These findings contribute to growing evidence that an enhanced intracellular pro-inflammatory response is a key mechanism through which social isolation may impact disease risk.

Background: The biological mechanisms underlying “chemo brain” or neurobehavioral symptoms in cancer patients are largely unknown, however, an emergent literature links indicators of immune deregulation to these symptoms. We hypothesized higher concentrations of pro-inflammatory cytokine interleukin 6 in breast cancer (BC) patients at both pre and post-treatment assessment relative to healthy controls, and that IL6 levels would be associated with changes in neurobehavioral functioning. Methods: Post-menopausal BC patients were assessed prior to any local or systemic treatment and one month after completion of primary treatment. Age-matched Healthy controls (HC) were assessed at similar time intervals. Neurocognitive, behavioral, and IL6 data for 120 BC and 56 HC were examined. Results: Statistically significant differences in IL6 levels between BC and HC were not evident at baseline but were at follow up assessment, with increased levels in the BC group. Factorial ANCOVA which evaluated pre-post treatment change in the BC and HC groups found that BC patients with higher IL6 levels at baseline had significant declines in selected cognitive functioning relative to the comparison groups, including memory retention ( F (1,155) = 7.07; p = .009) and processing speed ( p = .055). Similar interaction effects were not found on measures of fatigue, mood, or pain. Conclusion: Breast cancer patients with higher IL6 levels prior to treatment appear to be at risk for worse functioning in selected behavioral outcomes following treatment.

The inflammatory responses generated by partial sleep deprivation may have important health implications. Using 25 healthy adults (17F:8M), we examined both stimulated and unstimulated levels of several activated transcription factors involved in inflammatory responses (STAT1, STAT3, STAT5, and NFκB) via flow cytometry in peripheral blood subsets collected at baseline, after a night of partial sleep deprivation, and after a recovery night of sleep. In a repeated-measures ANOVA model, significant effects by time (p

IgE is an immunoglobulin isotype that is associated with allergy, and is produced in response to stimulation of B cells by the type 2 cytokines IL4 and IL13 and subsequent IgH class switch recombination and isotype switching. To determine the association between total IgE and risk of diffuse large B cell lymphoma (DLBCL) and follicular lymphoma, a nested case-control study was conducted among active-duty military personnel with archived serum in the US Department of Defense Serum Repository collected over several years prior to diagnosis of NHL. Cases were identified from the Armed Forces Institute of Pathology (AFIP) National Pathology Repository and the military Automated Centralized Tumor Registry (ACTUR). Each case was matched to two controls on age, sex, race, and dates of blood collection. Among study participants, the mean age at diagnosis was 44 years, 91% of the study subjects were male, and 66% were Caucasian. Because so few women were available for study, the analysis was restricted to men. IgE levels were quantified in one to three pre-NHL diagnosis serum specimens and in matched control specimens, using a human IgE ELISA that utilized the CIA-7.12 and CIA-4.15 monoclonal antibodies to IgE (0.67 ng IgE = 1 international unit (IU) as defined by WHO IgE standard NIBSC 75/502); the lower limit of detection in serum samples was 8 ng/ml. No significant differences were found in mean total IgE serum levels among men who went on to develop DLBCL (n=242, mean = 157 ng/ml; 95% CI: 135-180 ng/ml) compared with controls (n=480, mean = 150 ng/ml; 95% CI: 135-164 ng/ml). Similarly, null findings also were noted for men who developed follicular lymphoma (n=123, mean = 107 ng/ml; 95% CI: 82-131 ng/ml), compared with controls (n=244, mean = 120 ng/ml; 95% CI: 103-137 ng/ml). In contrast, mean serum IgE levels were significantly lower prior to multiple myeloma diagnosis (n=38, mean = 66 ng/ml; 95% CI: 50-82 ng/ml) compared with controls (n=75, mean = 136 ng/ml; 95% CI: 101-171 ng/ml) (p=0.006) among men in the same cohort. Using mixed-effects modeling, cases and controls showed no significant differences in total IgE serum levels across the time preceding DLBCL or follicular lymphoma diagnosis, controlling for age, race/ethnicity and time of blood draw. These results are consistent with the conclusion that there is no association of DLBCL or follicular lymphoma with pre-diagnosis total IgE levels. Others have reported that decreased pre-diagnosis levels of allergen-specific IgE were seen in those who developed NHL, or that total IgE levels were decreased post-NHL diagnosis. These data, obtained from pre-cancer diagnosis longitudinal specimens collected from presumably immunocompetent persons, should help elucidate the role of total IgE levels and risk of B cell malignancies. The views expressed are those of the authors and should not be construed to represent the positions of the Department of the Army or Department of Defense. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4644. doi:10.1158/1538-7445.AM2011-4644

Studies on several different types of carcinomas, with the notable exception of colon carcinoma, have shown that poorly differentiated tumors are frequently deficient in E-cadherin dependent cell-cell adhesion. In this study, we examined Ca(2+)-dependent cell-cell adhesion in colon carcinoma cell lines. Five poorly differentiated (Clone A, MIP 101, RKO, CCL 222, CCL 228) and four moderately-well differentiated (CX-1, CCL 235, DLD-2, CCL 187) colon carcinoma cell lines were assayed for their ability to form cell-cell aggregates and for their levels of E-cadherin expression. All of the poorly differentiated cell lines exhibited low levels of Ca(2+)-dependent cell-cell aggregation, in contrast to the moderately-well differentiated cell lines. Contrary to most previous studies, however, we observed that three of the five poorly differentiated cell lines examined expressed E-cadherin by FACS analysis and immunoprecipitation using an E-cadherin mAb. In fact, two of these cell lines expressed a 3- to 4-fold higher level of E-cadherin than that found in the moderately-well differentiated cell lines. mRNA levels for E-cadherin, as evaluated by both RT-PCR and Northern hybridization, corresponded to the levels of protein expression in each of the cell lines. Immunoprecipitation with an E-cadherin mAb, which is known to co-precipitate the catenins, demonstrated that the three poorly differentiated cell lines expressing E-cadherin did not co-precipitate alpha-catenin, although all of the moderately-well differentiated cell lines expressed both alpha- and beta-catenin. RT-PCR confirmed the absence of the alpha-catenin mRNA from two of these cell lines. Stable expression of an alpha-catenin cDNA in one of the poorly differentiated cell lines lacking alpha-catenin expression resulted in a 5-fold increase in its level of Ca(2+)-dependent cell-cell aggregation, providing evidence that alpha-catenin is directly responsible for the loss of cell-cell adhesion in some cell lines. The alpha-catenin transfectants also exhibited a marked reduction in migration on collagen I. These data indicate that loss of alpha-catenin expression, as well as E-cadherin expression, can lead to a phenotype associated with poorly differentiated colon carcinomas.

Background Multiple myeloma (MM) is a non-AIDS related malignancy of an immunoglobulin (Ig)-secreting B cell (plasma cell) usually associated with older age. MM is characterized by the presence of a high concentration monoclonal Ig protein in serum, and excretion of free light chains (FLC) in urine. In the years preceding MM diagnosis, a small monoclonal Ig develops that may be detectable upon serum protein electrophoresis, resulting in a premalignant condition known as monoclonal gammopathy of undetermined significance (MGUS). In an individual with MGUS, there may also be abnormal levels of FLC. Increased circulating levels of Ig, other indicators of B cell hyperactivation, and B cell lymphoma are well-recognized features of HIV infection. There have also been several reports of increased prevalence of MGUS and a meta-analysis showing increased risk of MM among HIV-infected (HIV+) individuals. There are, however, little or no longitudinal data on HIV-associated Ig abnormalities, B cell activation, and MGUS. We have initiated a pilot study within the Multicenter AIDS Cohort Study (MACS), a prospective study of untreated and treated HIV infection and AIDS in the United States, to begin to examine Ig abnormalities associated with MGUS in HIV-uninfected (HIV-) and HIV+ homosexual men.

Elevated serum levels of heat shock protein 70 precede the development of AIDS-non-Hodgkin lymphoma in carriers of the common and highly conserved HLA-B8-DR3 haplotype Brahim Aissani, Otoniel Martinez-Maza, Sadeep Shrestha, Elizabeth Breen, Lisa Jacobson, Richard Kaslow From 12 International Conference on Malignancies in AIDS and Other Acquired Immunodeficiencies (ICMAOI) Bethesda, MD, USA. 26-27 April, 2010

3-5 years: OR=3.66 (95% CI, 2.34-5.74); 1-3 years: OR=6.62 (95% CI, 3.78-11.6); and 0-1 year: OR=3.68 (95% CI, 2.27-5.98). Subgroup analyses revealed that CXCL13 was more strongly associated with systemic NHL compared to central nervous system NHL, and EBV-negative compared to EBV-positive tumors. Conclusions These data suggest that CXCL13 may be ab iomarker for NHL in the setting of HIV, and is more strongly associated with systemic, EBV-negative tumors. Studies are currently under way to further characterize the role CXCL13, and its receptor CXCR5, in lymphomagenesis.

We thank Dr. Krishnadas for his interest in our manuscript “Tumor Necrosis Factor Antagonism Normalizes Rapid Eye Movement Sleep in Alcohol Dependence” and his questions about the mechanisms that transduce peripheral blockade of proinflammatory cytokine activity (e.g., admininstration of an antagonist of tumor necrosis factor) to alterations in brain function and behavior (e.g., sleep). Although it was beyond the scope of this experimental, clinical trial to evaluate the pathways that signal changes in peripheral proinflammatory cytokines to changes in sleep in alcohol dependence, the mechanisms that might contribute to these behavioral changes have been extensively reviewed (1,2). As previously reviewed, basic research has implicated four relevant pathways that are thought to communicate peripheral proinflammatory cytokine activity to the brain. These pathways include the following: 1) in a neural pathway, local levels of proinflammatory cytokines activate primary afferent nerves, such as the vagal nerves; 2) in a humoral pathway, macrophage-like cells residing in the circumventricular organs and the choroid plexus produce proinflammatory cytokines, and these cytokines can enter the brain by volume diffusion; 3) in a cytokine transporter pathway, saturable transport systems at the blood–brain barrier facilitate pro-inflammatory cytokines overflowing in the systemic circulation to gain access to the brain through active transport; and 4) activation of IL□1 receptors that are located on perivascular macrophages and endothelial cells of brain venules results in the local production of prostaglandin E2. Together, these immune to immune-to-brain communication pathways ultimately leads to the production of proinflammatory cytokines by microglial cells, which in turn is thought to induce changes in brain function and behavioral responses. We agree that clinical translational studies are needed to understand the relevant pathways by which antagonism of peripheral cytokine activity leads to changes in sleep and possibly other behaviors (e.g., depressive symptoms) in humans (3)

Inflammation, part of the body's innate immune response, can lead to "sickness behaviors," as well as alterations in social and affective experiences. Elevated levels of pro-inflammatory cytokines have been associated with increased neural sensitivity to social rejection and social threat, but also decreased neural sensitivity to rewards. However, recent evidence suggests that inflammation may actually enhance sensitivity to certain social rewards, such as those that signal support and care. Despite a growing interest in how inflammation influences neural reactivity to positive and negative social experiences, no known studies have investigated these processes in the same participants, using a similar task. To examine this issue, 107 participants were randomly assigned to receive either placebo or low-dose endotoxin, which safely triggers an inflammatory response. When levels of pro-inflammatory cytokines were at their peak, participants were scanned using fMRI while they received positive, negative, and neutral feedback from an "evaluator" (actually a confederate) about how they came across in an audio-recorded interview. In response to negative feedback (vs. neutral), participants in the endotoxin condition showed heightened neural activity in a number of threat-related neural regions (i.e., bilateral amygdala, dorsal anterior cingulate cortex) and a key mentalizing-related region (i.e., dorsomedial PFC), compared to placebo participants. Interestingly, when receiving positive feedback (vs. neutral), endotoxin (vs. placebo) led to greater neural activity in the ventral striatum and ventromedial PFC, regions often implicated in processing reward, as well as greater activity in dorsomedial PFC. Together, these results reveal that individuals exposed to an inflammatory challenge are more "neurally sensitive" to both negative and positive social feedback, suggesting that inflammation may lead to a greater vigilance for both social threats and social rewards.

AIDS-associated Non-Hodgkin's lymphoma (AIDS-NHL) remains a problem even in the era of effective anti-retroviral therapy. Recent studies have suggested that the chemokine, CXCL13, and its receptor, CXCR5 may play a role in B cell tumors (non-AIDS-associated). Normally, CXCL13 is expressed in secondary lymphoid tissues and directs the homeostatic movement of CXCR5(+) B cells through these areas. To evaluate the role that CXCL13 and CXCR5 might play in AIDS-NHL, serum of individuals (n = 46) who ultimately developed AIDS-NHL was obtained from the Multicenter AIDS Cohort Study (MACS) at UCLA. The AIDS-NHL serum specimens tested were collected at a mean of 8.9 months prior to NHL diagnosis (SD = 7.9 months). Sera from AIDS (non-lymphoma), healthy HIV-positive, and HIV-negative control subjects were also included in the study. The mean CXCL13 level in the AIDS-NHL group (158 pg/ml, SD = 153) was ~50 percent higher than the AIDS control group (98.4 pg/ml, SD = 70.9, P = 0.02). Furthermore, CXCL13 levels correlated with sCD44 levels in the AIDS-NHL group (R = 0.31, P = 0.04), but not in the AIDS control group (R = 0.07; P = 0.7, data not shown); we previously showed that sCD44 levels are elevated prior to AIDS-NHL development. CXCL13 levels in the AIDS-NHL group were also ~2.5 times greater than levels in the HIV-positive group, and ~7 times greater than levels in the HIV-negative group (P < 0.001 for both comparisons). Next, tissue arrays were obtained from the AIDS & Cancer Specimen Resource (ACSR) that contained numerous sections of primary AIDS-NHLs, including both the Burkitt and diffuse large cell subtypes. By immunohistochemistry, all primary AIDS-NHLs (24/24) expressed CXCR5, and 22/24 of the AIDS-NHL specimens also showed expression of CXCL13. Cell lines derived from primary AIDS-NHL tumors also showed strong expression of CXCR5, and occasionally, low levels of expression of CXCL13. AIDS-NHL cell lines also demonstrated chemotaxis towards CXCL13. These results indicate that CXCL13 and CXCR5 may play a role in the biology of AIDS-NHL, possibly by affecting the movement of pre-malignant and/or malignant B cells.