The Ribosomal Protein S19 Suppresses Antitumor Immune Responses via the Complement C5a Receptor 1.

Relatively little is known about factors that initiate immunosuppression in tumors and act at the interface between tumor cells and host cells. In this article, we report novel immunosuppressive properties of the ribosomal protein S19 (RPS19), which is upregulated in human breast and ovarian cancer cells and released from apoptotic tumor cells, whereupon it interacts with the complement C5a receptor 1 expressed on tumor infiltrating myeloid-derived suppressor cells. This interaction promotes tumor growth by facilitating recruitment of these cells to tumors. RPS19 also induces the production of immunosuppressive cytokines, including TGF-ß, by myeloid-derived suppressor cells in tumor-draining lymph nodes, leading to T cell responses skewed toward Th2 phenotypes. RPS19 promotes generation of regulatory T cells while reducing infiltration of CD8+ T cells into tumors. Reducing RPS19 in tumor cells or blocking the C5a receptor 1-RPS19 interaction decreases RPS19-mediated immunosuppression, impairs tumor growth, and delays the development of tumors in a transgenic model of breast cancer. This work provides initial preclinical evidence for targeting RPS19 for anticancer therapy enhancing antitumor T cell responses.

Pulmonary alveolar macrophages contribute to the premetastatic niche by suppressing antitumor T cell responses in the lungs.

In contrast to tumor-associated macrophages, myeloid-derived suppressor cells, or inflammatory monocytes, functions of tissue resident macrophages, including alveolar macrophages (AM), in cancer were not well studied. Using a mouse model of breast cancer, we show that AM promote cancer metastasis to the lungs by suppressing antitumor T cells in this organ. AM accumulated in the premetastatic lungs through complement C5a receptor-mediated proliferation but not through recruitment from the circulation. AM preconditioned by breast tumors inhibited Th1 and favored generation of Th2 cells that had lower tumoricidal activity than Th1 cells. In addition, AM reduced the number and maturation of lung dendritic cells by regulating TGF-ß in the lung environment. Depletion of AM reversed immunosuppression imposed by these cells and strengthened local Th1 responses, which significantly reduced lung metastatic burden. C5a receptor deficiency, which also lessens myeloid-derived suppressor cells in the premetastatic niche, synergized with the depletion of AM in preventing metastasis, leading to protection of mice from lung metastases. This study identifies AM as a new component of the premetastatic niche, which is harnessed by tumors to impose immunosuppression, and as a new target for cancer immunotherapies to eliminate or reduce metastasis. Because the lungs are the most common target for hematogenous metastasis, this research offers a plausible explanation for susceptibility of the lungs to cancer metastasis.

Women's knowledge of abortion law and availability of services in Nepal.

This paper assesses women's awareness of the liberalization of abortion law and their knowledge of a place for obtaining abortion services in Nepal. The data are from the 2011 Nepal Demographic and Health Survey. The results are compared with data from a similar survey conducted in 2006. Variations in the two measures among several population sub-groups are analysed by performing logistic regression. Among women aged 15-44, 38.7% (CI: 37.8, 39.6) were aware of the legal status of abortion and 59.8% (CI: 58.9, 60.7) knew of a place to have an abortion. The percentages of both measures varied considerably by various population sub-groups. Over a 5-year period, knowledge of the legality of abortion increased by 6.4 percentage points, and awareness of service delivery sites increased by 3.3 percentage points. The increases in both measures were, however, largely limited to higher wealth quintiles and those with higher educational attainment. The results suggest the need to intensify efforts to educate women in Nepal, particularly the most disadvantaged women, about abortion law, including the conditions under which abortion is permitted, and where to access safe abortion services.

Cellular immune response in young children accounts for recurrent acute otitis media.

Acute otitis media (AOM) is a common disease in young children. Streptococcus pneumoniae (Spn) and Haemophilus influenzae (NTHi) are the two most common pathogens that cause AOM. Over the past 5 years, our group has been studying the immunologic profile of children that experience repeated AOM infections despite tympanocentesis drainage of middle ear fluid and individualized antibiotic treatment; we call these children stringently-defined otitis prone(sOP). Although protection against AOM is primarily mediated by ototpathogen-specific antibody, our recent studies suggest that suboptimal memory B and T cell responses and an immaturity in antigen-presenting cells may play a significant role in the propensity to recurrent AOM infections. This review focuses on the studies performed to define immunologic dysfunction in sOP children.

Reduced serum IgG responses to pneumococcal antigens in otitis-prone children may be due to poor memory B-cell generation.

A low level of serum antibody to antigens expressed by Streptococcus pneumoniae has been proposed to explain the susceptibility of children to recurrent episodes of acute otitis media (hereafter, "otitis-prone children"). By use of enzyme-linked immunospot assays, the percentages of memory B cells to pneumococcal protein antigens PhtD, LytB, PcpA, PhtE, and Ply were compared between otitis-prone and non-otitis-prone children at the time of acute otitis media or nasopharyngeal colonization with S. pneumoniae. We found significantly lower percentages of memory B cells to 3 pneumococcal protein antigens (PhtD, PhtE, and Ply) and reduced antigen-specific immunoglobulin G concentrations in otitis-prone children, compared with non-otitis-prone children.

Birth registration in Nepal: An assessment of progress based on two national surveys.

Birth registration, an essential component of the civil registration system, is expected to be complete and universal. This study assesses the progress made in recent years and identifies gaps in birth registration in Nepal. Data from the Multiple Indicator Cluster Surveys undertaken in 2014 and 2019 are used for the analysis. The two surveys included a total of 12,007 children under five years of age living with their mothers at the time of the surveys. The survey respondents were 11,821 mothers and 186 caretakers (in the case of those without mothers) of the children. The variations in the proportion of births registered among various subgroups of the children are assessed by performing bivariate analysis and binary logistic regression. Birth registration increased considerably, from 58% (95% CI: 57-59%) in 2014 to 77% (95% CI: 76-78%) in 2019. Several of the disparities between and among the various population subgroups that were evident in the 2014 survey had been considerably reduced or eliminated by 2019. The disparities in registration between boys and girls attenuated over time. Although birth registration increased for all children (ages 0-59 months old), infants still had comparatively lower levels of registration. The relatively disadvantaged provinces showed significant progress between the two survey periods. Considerable and significant progress has been made in birth registration in recent years. However, achieving universal and complete birth registration would require sustaining recent achievements and applying proven strategic interventions to ensure the inclusion of the unregistered births.

Reduced memory CD4+ T-cell generation in the circulation of young children may contribute to the otitis-prone condition.

BACKGROUND: An explanation for the immunologic dysfunction that causes children to be prone to repeated episodes of acute otitis media (AOM) has long been sought. Poor antibody response has been associated with the otitis-prone condition; however, there is no precise mechanistic explanation for this condition. METHODS: Non-otitis-prone and otitis-prone children with AOM or nasopharyngeal (NP) colonization caused by either Streptococcus pneumoniae or Haemophilus influenzae were compared for pathogen-specific CD4(+) T-helper memory responses by stimulating peripheral blood mononuclear cells using 6 vaccine candidate S. pneumoniae and 3 H. influenzae protein antigens. Samples were analyzed by multi-parameter flow cytometry. RESULTS: Significantly reduced percentages of functional CD45RA(Low) memory CD4(+) T cells producing specific cytokines (interferon γ, interleukin [IL]-2, IL-4 and IL-17a) were observed in otitis-prone children following AOM and NP colonization with either S. pneumoniae or H. influenzae. Immunoglobulin (Ig) G responses to the studied protein antigens were reduced, which suggests that antigen-specific B-cell function may be compromised as a result of poor T-cell help. Staphylococcal enterotoxin B stimulated similar cytokine patterns in memory CD4(+)T cells in both groups of children. CONCLUSIONS: Otitis-prone children have suboptimal circulating functional T-helper memory and reduced IgG responses to S. pneumoniae or H. influenzae after colonization and after AOM; this immune dysfunction causes susceptibility to recurrent AOM infections.

STimulator of INterferon Genes Agonism Accelerates Antitumor Activity in Poorly Immunogenic Tumors.

The innate immune agonist STING (STimulator of INterferon Genes) binds its natural ligand 2'3'-cGAMP (cyclic guanosine-adenosine monophosphate) and initiates type I IFN production. This promotes systemic antigen-specific CD8+ T-cell priming that eventually provides potent antitumor activity. To exploit this mechanism, we synthesized a novel STING agonist, MSA-1, that activates both mouse and human STING with higher in vitro potency than cGAMP. Following intratumoral administration of MSA-1 to a panel of syngeneic mouse tumors on immune-competent mice, cytokine upregulation and its exposure were detected in plasma, other tissues, injected tumors, and noninjected tumors. This was accompanied by effective antitumor activity. Mechanistic studies in immune-deficient mice suggested that antitumor activity of intratumorally dosed STING agonists is in part due to necrosis and/or innate immune responses such as TNF-α activity, but development of a robust adaptive antitumor immunity is necessary for complete tumor elimination. Combination with PD-1 blockade in anti-PD-1-resistant murine models showed that MSA-1 may synergize with checkpoint inhibitors but can also provide superior tumor control as a single agent. We show for the first time that potent cyclic dinucleotides can promote a rapid and stronger induction of the same genes eventually regulated by PD-1 blockade. This may have contributed to the relatively early tumor control observed with MSA-1. Taken together, these data strongly support the development of STING agonists as therapy for patients with aggressive tumors that are partially responsive or nonresponsive to single-agent anti-PD-1 treatment by enhancing the anti-PD-1 immune profile.

Increased sensitivity to antigen in high avidity CD8(+) T cells results from augmented membrane proximal T-cell receptor signal transduction.

The functional avidity of a cytotoxic T lymphocyte (CTL) is known to be a critical determinant of the efficacy with which it clears pathogens. High avidity cells, which are by definition highly sensitive to peptide antigen, are superior for elimination of viruses and tumours. Our studies have established the ability of T cells to undergo avidity modulation as a result of antigen encounter. High and low avidity cells established in this manner exhibit significant differences in the amount of peptide required to elicit effector function. However, how signalling is regulated in these cells as it relates to the control of peptide sensitivity remains to be defined. To address this question, we compared T-cell receptor (TCR) signal transduction events in high and low avidity CTL generated from OT-I(rag2-) TCR transgenic mice. Our data suggest that divergent signalling is initiated at the TCR-associated CD3ζ, with low avidity CTL requiring higher amounts of pMHC to achieve threshold levels of phosphorylated CD3ζ compared with high avidity CTL. Further, this difference is transduced further downstream to mitogen-activated protein kinase and Ca(2+) signalling pathways. These results suggest that regulated control of the initiation of TCR signalling in high versus low avidity cells determines the amount of peptide required for T-cell activation.

Evaluation of 5-aminoisoquinoline (5-AIQ), a novel PARP-1 inhibitor for genotoxicity potential in vitro and in vivo.

5-aminoisoquinoline (5-AIQ) is an active PARP-1 inhibitor as well as an important functional group various drugs. Quinolines are generally known as mutagenic and carcinogenic in various in vitro and in vivo systems, while both positive and negative findings are available on the mutagenic potential of several isoquinolines. Since no literature is available on the genotoxicity of 5-AIQ, a battery of tests were conducted, in accordance with relevant OECD protocols, such as bacterial reverse mutation test, in vitro chromosomal aberration test, and bone marrow micronucleus test in mouse. These studies demonstrate that 5-AIQ does not possess genotoxic activity both with in vitro and in vivo systems. The findings substantiate the therapeutic value of 5-AIQ.

Use of liposomized tetracycline in elimination of Wolbachia endobacterium of human lymphatic filariid Brugia malayi in a rodent model.

Wolbachia bacteria, being filarial parasite symbiont have been implicated in a variety of roles, including development, fecundity and the pathogenesis of the filarial infections. Among various strategies used in the treatment of experimental filariasis, the elimination of symbiont Wolbachia seem to offer an efficient means of curing the disease. The antiwolbachial property of tetracycline has been well worked out; however, treatment needs to be continued for a prolonged period of time to achieve complete elimination of Wolbachia from the filarial parasites and their subsequent killing. This results in acute toxicity, thus limiting its practical utility for clinical implementation. In order to increase efficacy of the antibiotic with minimal toxic manifestations, we developed liposomized formulation of the tetracycline. The liposomized tetracycline was found to be significantly more effective when compared to the free form of the drug. In contrast to the 90/120 days oral administration of the drug, the treatment schedule using the liposomized form of the drug was reduced to 12 alternate days with better efficacy of the treatment.

Women's awareness of liberalization of abortion law and knowledge of place for obtaining services in Nepal.

In Nepal, following the liberalization of the abortion law, expansion and scaling up of services proceeded in parallel with efforts to create awareness of the legalization status of abortion and provide women with information about where services are available. This article assesses the effectiveness of these programmatic interventions in the early years of the country's abortion program. Data from a 2006 national survey are analyzed with 2 outcome measures-awareness of the legal status of abortion and knowledge of places to obtain abortion services among women ages 15 to 44 years. The variations in the outcomes are analyzed by ecological-development subregion, residence, education, household wealth quintile, age, and number of living children. Bivariate and multivariate logistic regression techniques are used. Overall 32.3% (95% confidence interval = 31.4% to 33.2%) of the respondents were aware of the legal status of abortion and 56.5% (95% confidence interval = 55.5% to 57.4%) knew of a place where they could obtain an abortion. Both outcome measures showed considerable variations by the covariates. Women with secondary or higher level of education had the highest odds ratio of being aware of the law and having knowledge of a source for abortion services. Ecological-development subregions showed the second highest levels of odds ratios. Significant disparities among the population subgroups existed in the diffusion of awareness of the legal status of abortion and having knowledge of a place for abortion services in Nepal. The results point to which population subgroups to focus on and also serve as a baseline for assessing future progress in the diffusion process.

Complement c5a receptor facilitates cancer metastasis by altering T-cell responses in the metastatic niche.

The impact of complement on cancer metastasis has not been well studied. In this report, we demonstrate in a preclinical mouse model of breast cancer that the complement anaphylatoxin C5a receptor (C5aR) facilitates metastasis by suppressing effector CD8(+) and CD4(+) T-cell responses in the lungs. Mechanisms of this suppression involve recruitment of immature myeloid cells to the lungs and regulation of TGFß and IL10 production in these cells. TGFß and IL10 favored generation of T regulatory cells (Treg) and Th2-oriented responses that rendered CD8(+) T cells dysfunctional. Importantly, pharmacologic blockade of C5aR or its genetic ablation in C5aR-deficient mice were sufficient to reduce lung metastases. Depletion of CD8(+) T cells abolished this beneficial effect, suggesting that CD8(+) T cells were responsible for the effects of C5aR inhibition. In contrast to previous findings, we observed that C5aR signaling promoted Treg generation and suppressed T-cell responses in organs where metastases arose. Overall, our findings indicated that the immunomodulatory functions of C5aR are highly context dependent. Furthermore, they offered proof-of-concept for complement-based immunotherapies to prevent or reduce cancer metastasis.

Deficiencies in the CD4+ T-Helper Cell Arm of the Immune System of Neonates and Young Children.

Newborns and young children rely on innate immunity to protect against infections until the adaptive immune system matures. Immunization helps facilitate protection, but multiple doses are needed to establish sufficient antibody levels and T-cell-facilitated immune memory. Deficient T-cell activation and function among neonates and young children are primarily present in the CD4+ compartment, whereas CD8+ T-cell function is at par with adults. CD4+ T cells in neonates and young children produce low levels of IFNγ, interleukin (IL)-2, IL-13, IL-5, and IL-17. This inherent deficiency in neonatal and young child CD4+ T-cell functionality has been linked to several mechanistic failures: (1) lower sensitivity to T-cell receptor stimulation, (2) increased apoptosis after proliferation, (3) unavailability of antigen for T-cell priming, and (4) inefficient stimulation by relatively immature antigen-presenting cells. In this review, we discuss evidence from infection and vaccination responses that shed light on the various checkpoints possibly involved in delayed maturation of CD4+ T-cell activation and function in newborns and young children.

CD4+ T-cell responses among adults and young children in response to Streptococcus pneumoniae and Haemophilus influenzae vaccine candidate protein antigens.

We characterized cytokine profiles of CD4(+) T-helper (h) cells in adults and young children to ascertain if responses occur to next-generation candidate vaccine antigens PspA, PcpA, PhtD, PhtE, Ply, LytB of Streptococcus pneumonia (Spn) and protein D and OMP26 of non-typeable Haemophilus influenzae (NTHi). Adults had vaccine antigen-specific Th1 and Th2 cells responsive to all antigens evaluated whereas young children had significant numbers of vaccine antigen-specific CD4(+) T cells producing IL-2, (p=0.004). Vaccine antigen-specific CD4(+) T-cell populations in adults were largely of effector (TEM) and/or central memory (TCM) phenotypes as defined by CD45RA(-)CCR7(+) or CD45RA(-)CCR7(-) respectively; however among young children antigen-specific IL-2 producing CD4(+) T cells demonstrated CD45RA(+) expression (non-memory cells). We conclude that adults have circulating memory CD4(+) T cells (CD45RA(-)) that can be stimulated by all the tested Spn and NTHi protein vaccine candidate antigens, whereas young children have a more limited response.

PcpA of Streptococcus pneumoniae mediates adherence to nasopharyngeal and lung epithelial cells and elicits functional antibodies in humans.

Streptococcus pneumoniae (pneumococci) adhere to human nasopharyngeal (NP) epithelial cells as a first step in colonization and adherence of pneumococci to lung epithelia may be required to establish pneumonia. We sought to determine if PcpA can serve as an adhesin to human NP (D562) and lung (A549) epithelial cells and whether PcpA mediated adherence can be inhibited by human anti-PcpA antibodies. A PcpA isogenic mutant was constructed in a pneumococcal TIGR4 background. When the mutant and wild type strains were compared for their adherence to D562 and A549 cell lines, a reduction in adherence by the mutant was observed (p = 0.0001 for both cell types). PcpA was ectopically expressed on the surface of minimally-adherent heterologous host Escherichia coli resulting in augmented adherence to D562 (p = 0.002) and A549 (p = 0.015) cells. Total IgG was purified from a pool of 6 human sera having high IgG titers of anti-pneumococcal proteins. The purified IgG reduced TIGR4 adherence to D562 cells but we determined that this effect was largely due to bacterial cell aggregation as determined by flow cytometry and confocal microscopy. Fab fragments were prepared from pooled IgG sera. Inhibition of TIGR4 adherence to D562 cells was observed using the Fab fragments without causing bacterial aggregation (p = 0.0001). Depletion of PcpA-specific Fab fragments resulted in an increase in adherence of TIGR4 to D562 cells (p = 0.028). We conclude that PcpA can mediate adherence of pneumococci to human NP and lung epithelial cells and PcpA mediated adherence can be inhibited by human anti-PcpA antibodies.

Pivotal Advance: Nonfunctional lung effectors exhibit decreased calcium mobilization associated with reduced expression of ORAI1.

CD8(+) T cells play a critical role in the clearance of respiratory pathogens. Thus, it is surprising that functional inactivation of lung effectors has been observed in many models of viral infection. Currently, the molecular defect responsible for the shut-off of function in these cells is unknown. In the present study, we addressed this question using a model of respiratory infection with the paramyxovirus SV5. Nonfunctional cells were found to exhibit decreases in SOCE, resulting in reduced NFAT1 activation. Notably, function could be restored by the provision of increased levels of extracellular calcium. The reduced ability to mobilize calcium was associated with reduced expression of ORAI1, the CRAC channel subunit. These findings reveal a previously unknown mechanism for the negative regulation of function in effector T cells.

Patterns of smoking among adolescents in Malaysia and Thailand: findings from the International Tobacco Control Southeast Asia survey.

At present, 70% of the world's 1.1 billion smokers are in developing countries, with over 50% in Asia alone. The current study examined patterns of youth smoking in Thailand and Malaysia. Respondents were 2002 youths between the ages of 13 and 17 from Thailand (n = 1000) and Malaysia (n = 1002). Respondents were selected using a multistage cluster sampling design and surveyed between January 2005 and March 2005. Approximately 3% of youth between the ages of 13 and 17 were current smokers, with an additional 10% to 12% reporting experimental smoking. Males were between 7 and 15 times more likely to report smoking behavior than females. Less than 1% of females respondents in either country met the criteria for current smoking, and less than 5% met the criteria for experimental smoking. In contrast, more than 50% Thai males and approximately one-third of Malaysian males aged 17 met the criteria for either experimental or current smoking.