Role of inflammatory mediators in resistance and susceptibility to pneumococcal infection

Variations in the host response during pneumonia caused by Streptococcus pneumoniae in susceptible (CBA/Ca) and resistant (BALB/c) inbred mouse strains were investigated. Significant differences were detected in survival time, core body temperature, lung-associated and systemic bacterial loads, mast cell numbers, magnitude and location of cytokine production, lung disruption, and ability of isolated lung cells to release the cytokine tumor necrosis factor (TNF) alpha in vitro. Overall, the results indicate that the reduced capacity of CBA/Ca mice to induce rapid TNF activity within the airways following infection with S. pneumoniae may be a factor in their elevated susceptibility to pneumococcal pneumonia.     

Differential susceptibility of inbred mouse strains to Burkholderia thailandensis aerosol infection

Burkholderia pseudomallei is the causative agent of melioidosis, an emerging bacterial disease that accounts for high rates of septicaemia and death in parts of Southeast Asia and Northern Australia. The closely related species Burkholderia thailandensis is considered avirulent in humans and has been used as a surrogate for B. pseudomallei in several studies. The pathogenesis of B. pseudomallei and the role of Toll-like receptors (TLRs) in host immunity to infection are not well-defined. In this study, we exposed four strains of inbred mice (BALB/c, C57BL/6, TLR4-deficient C3H/HeJ, and TLR4-competent C3H/HeN) to increasing doses of aerosolized B. thailandensis to determine strain susceptibility and the role of TLR4 during pulmonary infection. Our results indicate an increased susceptibility in the C57BL/6 and BALB/c strains, who displayed lethality, bacterial burden in organs, and pulmonary and systemic inflammation. C3H/HeJ were as resistant as C3H/HeN mice to B. thailandensis at the highest challenge dose examined, but TLR4-deficient animals exhibited a modest increase in chronic pulmonary inflammation. These results demonstrate that B. thailandensis can be used as a surrogate for experimental laboratory investigation of melioidosis in small animal models and that TLR4 may not play a prominent role during acute pneumonic melioidosis.     

Forward genetic dissection of innate response to infection in inbred mouse strains: selected success stories

Mouse genetics is a powerful tool for the dissection of genes, proteins, and pathways important in biological processes. Application of this approach to study the host response to infection has been a rich source of discoveries that have increased our understanding of the early innate pathways involved in responding to microbial infections. Here we review some of the key discoveries that have arisen from pinpointing the genetic defect in mouse strains with unusual or extreme response to infection and have led to insights into pathogen sensing pathways and downstream effector functions of the early innate immune response.     

Genetic control of susceptibility to infection with Plasmodium chabaudi chabaudi AS in inbred mouse strains

To identify genetic effects modulating the blood stage replication of the malarial parasite, we phenotyped a group of 25 inbred mouse strains for susceptibility to Plasmodium chabaudi chabaudi AS infection (peak parasitemia, survival). A broad spectrum of responses was observed, with strains such as C57BL/6J being the most resistant (low parasitemia, 100% survival) and strains such as NZW/LacJ and C3HeB/FeJ being extremely susceptible (very high parasitemia and uniform lethality). A number of strains showed intermediate phenotypes and gender-specific effects, suggestive of rich genetic diversity in response to malaria in inbred strains. An F2 progeny was generated from SM/J (susceptible) and C57BL/6J (resistant) parental strains, and was phenotyped for susceptibility to P. chabaudi chabaudi AS. A whole-genome scan in these animals identified the Char1 locus (LOD=7.40) on chromosome 9 as a key regulator of parasite density and pointed to a conserved 0.4-Mb haplotype at Char1 that segregates with susceptibility/resistance to infection. In addition, a second locus was detected in [SM/J × C57BL/6J] F2 mice on the X chromosome (LOD=4.26), which was given the temporary designation Char11. These studies identify a conserved role of Char1 in regulating response to malaria in inbred mouse strains, and provide a prioritized 0.4-Mb interval for the search of positional candidates.     

Experimental Borrelia burgdorferi infection in inbred mouse strains: antibody response and association of H-2 genes with resistance and susceptibility to development of arthritis

We have investigated the specific humoral immune response and its correlation to the development of disease after experimental inoculation of B. burgdorferi in different inbred strains of mice. All mouse strains tested showed high levels of specific IgM antibodies during the initial 10 days of infection. Specific IgG antibodies predominantly of the IgG2a, IgG2b and IgG3 isotypes were found in increasing amounts by 14 days post infection. Antibody titers peaked at days 65 and 110. Particularly low titers of specific IgM and/or IgG antibodies were detected in sera of AKR/N and B10.BR mice. Antibodies specific for numerous B. burgdorferi antigens including the outer surface proteins A (31 kDa) and B (34 kDa) and a protein(s) of molecular mass of approximately 40 kDa, most probably 41 kDa (flagellin) and/or 39 kDa (p39), were induced in all inbred mouse strains within 2 weeks inoculation albeit in varying concentrations. Later during infection, the patterns of antibody specificities were much more complex. With regard to development of disease all strains of mice tested fall into three groups: (a) mice of H-2k haplotype (AKR/N, C3H/HeJ, C3H/HeN, B10.BR) developed a chronic progressive arthritis in the tibiotarsal joints, (b) mice of H-2 haplotypes, H-2b (C57BL/6), H-2j (B10.WB), H-2r (B10.R111) and H-2s (B10.S) developed arthritis of variable duration and intensity which was not progressive and (c) mice of H-2d haplotype (BALB/c, DBA/2, C.B-17, B10.D2, Cal.20), irrespective of their background genes or Igh allotype, showed no clinical signs of arthritis at any time point following inoculation of B. burgdorferi organisms. The finding of similar patterns of apparently protective antibodies in all mouse strains tested together with the striking association between the H-2d haplotype and resistance, and between the H-2k haplotype and the occurrence of B. burgdorferi-induced arthritis suggest a critical role of T cells in the development of the disease in mice.     

宿主的遗传背景对呼吸道感染沙眼衣原体后Treg产生的影响

目的 探讨宿主的遗传背景对呼吸道感染沙眼衣原体后调节性T细胞(Treg)产生的影响.方法 对衣原体感染具有明显易感性差异的C57 BL/6(C57)和C3H/HeN(C3H)小鼠鼻腔吸入1×103 IFU沙眼衣原体小鼠肺炎菌株(Chlamydia muridarum,Cm),于感染后不同天数处死小鼠.利用细胞内细胞因子染色技术检测小鼠脾脏单个核细胞CD4+ CD25+T细胞、Foxp3+ CD4+ CD25+T细胞百分率,利用RT-PCR技术检测小鼠肺组织Treg细胞分泌的相关细胞因子IL-10和IL-2的mRNA表达水平,并比较Cm呼吸道感染不同时期C57和C3H小鼠Treg免疫应答水平的差异.结果 Cm感染在两组小鼠均诱导较高水平的CD4+ CD25+T细胞、Foxp3+ CD4+ CD25+T细胞产生及IL-10、IL-2mRNA表达.感染后第3天和第7天,高易感性的C3H小鼠脾脏CD4+ CD25+T细胞、Foxp3+ CD4+CD25+T细胞扩增水平,以及肺组织细胞因子IL-2 mRNA的表达水平均高于C57小鼠,感染后第14天,C3H小鼠IL-10 mRNA表达水平明显高于C57小鼠.结论 衣原体呼吸道感染在高易感性的C3H小鼠诱导高水平的Treg的增殖及Treg相关细胞因子IL-10、IL-2的表达,从而对衣原体特异的Th1免疫应答抑制作用增强,在小鼠衣原体呼吸道感染易感性差异中发挥重要作用.     

Natural resistance to Salmonella typhimurium in different inbred mouse strains.

The mechanisms of natural resistance to intravenous challenge with Salmonella typhimurium C5 are complex. LD50 determinations showed inbred mouse strains of low, intermediate and high natural resistance, with BALB/c and B10 strains the most susceptible, A/J the most resistant. Delayed (footpad) hypersensitivity was not by itself a measure of natural resistance. Resistant mouse strains sensitized either s.c. or i.v. with an attenuated salmonella strain showed positive 48 h footpad reactions when tested 8 days later with a salmonella extract, but three very susceptible strains also showed positive reactions. Determinations of the in vivo net growth rate of salmonellae in the liver and spleen during the first phase of the infection (up to day 4) arrange the different mouse strains into two categories of fast and slow net growth rate. All fast net growth rate strains are susceptible, but not all slow net growth rate strains are resistant. Besides slow net growth rate, resistance requires the participation of other factors appearing in the second phase of the infection (towards the end of the first week) probably involving the cellular immune response, which halts further bacterial growth. Not all slow net growth rate strains are equally capable of suppressing bacterial growth in this second phase. The host mechanism determining slow net growth rate is inherited as a dominant trait, and appears to be operating before the main cellular immune response. The influence of this mechanism on net growth rate is reflected in the time to death following a given dose of salmonellae. The present results suggest that overall resistance to salmonellae is polygenic, but that the mechanism responsible for the differences in early net growth rate is less complex.     

Role of the host in pathogenesis of Helicobacter-associated gastritis: H. felis infection of inbred and congenic mouse strains.

In humans, Helicobacter pylori establishes a chronic infection which can result in various degrees of gastric inflammation, peptic ulcer disease, and a predisposition to gastric cancer. It has been suggested that bacterial virulence factors such as the vacuolating toxin (VacA) and the cytotoxin-associated gene product (CagA) may play a major role in determining the clinical outcome of Helicobacter infections. The role of host responses in these varied outcomes has received little attention. Helicobacter felis, which does not express CagA or VacA, causes chronic infection and inflammation in a well-characterized mouse model. We have used this model to evaluate the role of host responses in Helicobacter infections. BALB/c, C3H, and C57BL/6 mice were orally infected with a single strain of H. felis, and 2 and 11 weeks after infection, the mice were sacrificed and evaluated histologically for magnitude of H. felis infection. Intensity and extent of inflammation, and cellular composition of the inflammatory infiltrate. All three strains of mice demonstrated comparable levels of infection at 11 weeks, but the pattern and intensity of inflammation varied from minimal in BALB/c mice to severe in C57BL/6 mice. Gastric epithelial erosions were noted in C3H mice, and mucous cell hyperplasia was observed in C3H and C57BL/6 mice. Abundant mucosal mast cells were observed in the gastric tissues of all three mouse strains. Studies using major histocompatibility complex (MHC)-congenic mice revealed probable contributions by both MHC and non-MHC genes to Helicobacter-induced inflammation. Thus, large variations in the severity of disease were observed after infection of different inbred strains and congenic mice with a single isolate of H. felis. These results demonstrate the importance of the host response in disease outcome following gastric Helicobacter infection.     

Early gene expression differences in inbred mouse strains with susceptibility to pulmonary adenomas

Lung cancer is the most common cause of cancer-related deaths in both men and women, and effective preventatives are rare due to the difficulty of early detection. Specific gene expression signatures have been identified in individuals that already developed lung cancer. To identify if gene expression differences could be detected in individuals before the onset of the disease, we obtained lung tissues for microarray analysis from young, healthy mice of 9 inbred strains with known differences in their susceptibility to spontaneous pulmonary adenomas when aged. We found that the most common differentially expressed genes among all possible 36 strain comparisons showed significant associations with cancer- and inflammation-related processes. Significant expression differences between susceptible and resistant strains were detected for Aldh3a1, Cxcr1 and 7, Dpt, and Nptx1—genes with known cancer-related functions, and Cd209, Cxcr1 and 7, and Plag2g1b—genes with known inflammatory-related functions. Whereas Aldh3a1, Cd209, Dpt, and Pla2g1b had increased expression, Cxcr1 and 7, and Nptx1 had decreased expression in strains susceptible to pulmonary adenomas. Thus, our study shows that expression differences between susceptible and resistant strains can be detected in young and healthy mice without manifestation of pulmonary adenomas and, thus, may provide an opportunity of early detection. Finally, the identified genes have previously been reported for human non-small cell lung cancer suggesting that molecular pathways may be shared between these two cancer types.     

Direct genetic and maternal influences on behavior and growth in two inbred mouse strains

Newborn male DBA/2 and C57BL/6 mice were either crossfostered, fostered within strains, or, after handling, left with their own dams and compared for nine behavioral components observed in single animals at an adult age and for body weight measured at 4 weeks and 3 months. Many strain differences between the unfostered control groups emerged and these remained largely unchanged in comparisons between the infostered and between the outfostered groups. In neither infostered group were significant influences of fostering per se found. Some variation due to foster strain was detected mainly in the DBA outfostered group, namely, for 4-week body weight, rearing frequency, and locomotor activity, but no resemblance to the alien strain was brought about by this. Effects of litter occurred rarely. Unlike the C57BL strain, the DBAs showed a clear pattern of interdependence among exploratory acts. In spite of problems that may be attached to the crossfostering method, it was concluded that postnatal genotype-dependent maternal influences were rather unimportant as a source of variation in behavior and growth as measured between these two strains, relative to direct genetic effects and, possibly, prenatal factors.     

Characterization of chronic bronchopulmonary Pseudomonas aeruginosa infection in resistant and susceptible inbred mouse strains

Chronic bronchopulmonary Pseudomonas aeruginosa infection, initiated by intratracheal instillation of 1 to 2 x 10(5) colony-forming units of a mucoid strain of bacteria trapped in agar beads, was characterized in resistant BALB/c mice and susceptible C57BL/6 (B6) mice through 28 d postinfection. B6 mice experienced a more severe infection than BALB/c mice as evidenced by significantly higher mortality and significantly greater weight loss during the first 14 d. Furthermore, B6 mice had significantly higher numbers of bacteria in the lungs through 21 d after infection. Overall, only 22% of these hosts cleared the infection. In contrast, 67% of BALB/c mice cleared the infection. These differences between resistant and susceptible mice were found to correlate with histopathologic differences in the type of inflammation and the extent of tissue damage. An acute, predominantly neutrophilic inflammation and extensive tissue damage were apparent in the lungs of susceptible B6 mice, whereas chronic, granulomatous inflammation and little or no tissue damage were visible in resistant BALB/c mice. The finding of acute inflammation in the lungs of infected B6 mice was confirmed by fluorescence-activated cell sorter (FACS) analyses, which demonstrated that these mice had significantly greater proportions of polymorphonuclear neutrophils in the lungs on Days 7 and 14 after infection than did BALB/c mice. FACS analyses also revealed significant and similar increases in CD3(+) lung cells in both strains as the infection progressed. The CD4/CD8 ratio was significantly greater in BALB/c mice by 21 d after infection when the majority of these animals, but not B6 mice, had cleared the infection.     

Neuroendocrine responses regulating susceptibility and resistance to autoimmune/inflammatory disease in inbred rat strains

Summary: Rodent animal models of inflammatory and autoimmune disease have been important tools in the study of the interaction between neuroendocrine physiology and the immune responses. The rat has been particularly useful in part because, in contrast to other species, most rat models of autoimmune/inflammatory disease are induced rather than spontaneous. This allows for systematic and controlled manipulations of the neuroendocrine system in relation to exposure to the antigen or pro-inflammatory trigger. The most frequently used immune challenges include lipopolysaccharide-induced septic shock, carrageenan-induced local inflammation and adjuvant or bacterial cell wall-induced arthritis. By analyzing the responses to these challenges in different strains of rats and mice it has been possible to define the relationships between the neuroendocrine and immune systems and to identify some mechanisms through which these connections confer susceptibility and resistance to autoimmune and inflammatory diseases. The present review will discuss data obtained from rodent physiology, indicating that an important component in the susceptibility or resistance to development of these diseases is due to dysfunctional regulation of the immune response by the neuroendocrine hypothalamic–pituitary–adrenal axis. In particular, the importance of neurons of the paraventricular hypothalamic nucleus in determining susceptibility or resistance to autoimmune and inflammatory disease will be discussed.     

The midbrain dopaminergic system: anatomy and genetic variation in dopamine neuron number of inbred mouse strains

The mesotelencephalic dopamine system is genetically variable and affects motor behavior, motivation, and learning. Here we examine the genetic variation of mesencephalic DA neuron number in a quasi-congenic RQI mouse strain and its background partner and in a recombinant inbred strain with different levels of mesencephalic tyrosine hydroxylase activity (TH/MES). We used B6.Cb4i5-6/Vad, C57BL/6By, and CXBI, which are known to express high, intermediate, and low levels of TH/MES, respectively. Unbiased stereological sampling with optical disector counting methods were employed to estimate the number of TH-positive neurons in the A8-A9-A10 cell groups. Morphometric studies on the mesencephalic dopamine cell groups indicated that male mice of the B6.Cb4i5-6/Vad strain were endowed with a significantly lower number of TH-positive cells than CXBI mice. In all strains studied, the right retrorubral field (A8 area) had a higher number of dopamine neurons compared to the left A8 area. The results suggest an inverse relationship between TH/MES and number of dopamine neurons in the A9-A10 cell groups and significant lateral asymmetry in the A8 cell group. A detailed anatomical atlas of the mesencephalic A8-A9-A10 dopaminergic cell groups in the mouse is also presented to facilitate the assignment of TH-positive neurons to specific cell groups.     

Differences in resistance to Trypanosoma musculi infection among strains of inbred mice.

Inbred strains of mice were inoculated with Trypanosoma musculi, and the course of the ensuing parasitemia was followed. The mouse strains fell into three groups: those displaying high and moderate (fivefold less) parasitemia and C57BL/6 (B/6) mice which had exceptionally low infections. To gain insight concerning the mechanisms responsible for interstrain variations in infections, several types of experiments were performed. Comparison of the ability of spleen cells from the various strains to provide the growth-promoting substances required by T. musculi for growth in culture revealed that B/6 cells were deficient; this suggested one mechanism for regulating parasite infections. Exposure of C3H (high parasitemia) and B/6 mice to graded levels of ionizing radiation revealed that B/6 mice have much greater innate resistance to infection than do C3H mice. The effects of treating mice with silica dust or mercaptoethanol indicated that relative resistance to infection is not primarily associated with macrophage activity or limited growth-promoting substances. We conclude that variations in immune responsiveness to parasite antigens (probably not associated with the H-2 complex), possibly in concert with variations in a non-immunological mechanism, account for interstrain variation in resistance to T. musculi infections.     

Widespread susceptibility among inbred mouse strains to the induction of lupus autoantibodies by pristane

Unlike other agents associated with drug-induced lupus, the isoprenoid alkane pristane induces autoantibodies pathognomonic of lupus, including anti-Sm, anti-dsDNA, and anti-ribosomal P in BALB/c and SJL/J mice. The susceptibility of other strains of mice to pristane-induced lupus is unknown and is the focus of the present study. Anti-nRNP/Sm, anti-Su, and anti-ribosomal P autoantibodies were produced by most strains of mice surveyed within several months of pristane treatment, although there was marked interstrain variability in their frequencies, levels, and times of onset. In sharp contrast, the production of autoantibodies against the double-stranded RNA binding proteins NF45/NF90/p110 was restricted to B6 and B10.S mice. We conclude that pristane selectively induces lupus-specific autoantibodies in virtually any strain of mouse regardless of its genetic background. However, H-2-linked as well as non-H2 genes influenced the expression of individual autoantibody markers. The widespread susceptibility of pristane-treated mice to lupus autoantibody production and the relatively small effect of MHC are unique features of this chemically induced lupus syndrome, with potential implications for understanding the pathogenesis of autoantibodies in idiopathic human systemic lupus erythematosus.     

Epidemiology of invasive pneumococcal infection in Taiwan: antibiotic resistance,serogroup distribution,and ribotypes analyses

From July, 1998, to June, 1999, pneumococcal isolates from 288 patients with invasive disease in Taiwan were serogrouped and tested for their susceptibility to various antibiotics. Automated ribotyping was used to study their molecular epidemiology. The mortality rate among those > or = 65 years was higher than those 18 or 19-64 years (p < 0.001). The total incidence of infection was significantly higher during the cooler season than the warmer season (p = 0.017). Among strains isolated from children aged < or = 18 years, 76% were not susceptible to penicillin, a rate that was significantly higher (p < 0.001) than that for adults (45%), as was the susceptibility to azithromycin, erythromycin, and trimethoprim-sulfamethoxazole (p < 0.005). The most prevalent serogroup encountered in the invasive isolates was 23, followed by 6, 14, 19, and 3. Isolation of Streptococcus pneumoniae in cerebrospinal fluid was at high rate in children under 5 years (p = 0.00012). Molecular typing revealed a high degree of polymorphism among the isolates. Among serogroup 23 and 19 isolates, a high proportion had the same ribotypes, the Taiwan23F-15 and Taiwan19F-14 isolates, suggesting the circulation of a Taiwanese epidemic strain. In Taiwan, S. pneumoniae isolates should be tested for their resistance profile for children < or = 18 years old, as these are more likely to harbor high-level resistance. Control of pneumococcal infection with the 7-valent-conjugated vaccine should also be considered because it is estimated that it would cover nearly 90% of the serotypes among pediatric invasive disease.     

Mannose-binding lectin genotype and invasive pneumococcal infection

Invasive pneumococcal disease is a serious infection that primarily affects young children and elderly or immunocompromised persons, but it also can affect healthy persons. Mannose-binding lectin (MBL) is a mediator of innate host immunity that activates the complement pathway and directly opsonizes pathogens. Variant structural codon and promoter MBL alleles have been associated with susceptibility to infections. Sixty-three Belgian patients with invasive pneumococcal disease and 162 healthy Belgian controls were genotyped for MBL alleles. We found a nonsignificant increased risk between the MBL structural codon variants (52, 54, and 57) and invasive pneumococcal disease. Combining our data with similar data from Kronberg et al. (J Infect Dis 2002;185:1517-20) indicated that MBL structural variants contributed to a small but significant increased risk of invasive pneumococcal disease. On the other hand, the -221 and -550 promoter allele distribution and the prevalence of the combined MBL structural and promoter -221 variant alleles were not significantly different between the patient group and the control group.