Infections due to the Gram-negative bacteria Neisseria meningitidis and N. gonorrhoeae represent major public health problems around the world. Meningococcal infections total 0.5-1.2 million and cause death of 50,000-135,000 individuals annually worldwide. Infections by N. meningitidis can present with a range of symptoms including sudden onset of fever, headache, stiff neck, nausea, and alterations in mental state. The bacteria primarily infect the central nervous system causing cerebrospinal meningitis, but can also cause disseminated disease leading to an overwhelming inflammatory response known as sepsis that can result in vascular leakage, failure in multiple organs, and death or long term sequelae including amputation of limbs, deafness, and seizures. Declines in meningococcal disease have occurred in the last decade in many developed countries due in part to use of polysaccharide-protein conjugate vaccines to specific serogroups of N. meningitidis, although the vaccines are not 100% effective and do not provide protection against all serogroups or strains. In addition, two new vaccines for serogroup B disease have been approved recently, however approval is only for persons aged 10-25 years, and unanswered questions exist regarding efficacy. Paradoxically, N. meningitidis infects the nasopharynx of 8-20% of the population without causing disease. There are an estimated 106 million new cases of sexually transmitted gonococcal infections worldwide each year. Those most affected by the disease are women in whom infections are often asymptomatic. From 10- 20% of infected women suffer from pelvic inflammatory disease that can cause chronic pain, infertility, and ectopic pregnancy. Importantly, a growing number of studies have shown that gonococcal infection can facilitate the transmission of HIV. The lack of a vaccine and increasing number of strains of N. gonorrhoeae that are resistant to treatment with antibiotics have heightened the possibility of loss of control of the transmission of gonorrhea. Our work has focused on understanding the interactions of a major component of the bacterial membrane, the lipooligosaccharide (LOS), with the human immune system. Preformed components of what is termed the innate immune system are the first line of defense in protection against Gram-negative bacteria such as Neisseria. We have found that the ability of Neisseria to induce a feedback down-regulation of the innate immune response is positively correlated with the inflammatory potential of the bacteria which in turn is mediated by the expression of particular molecular substituents on the LOS. Our data also show that certain elements of the LOS structure assist the bacteria in evading lysis by the complement cascade in the blood and recognition by human antibodies that facilitate phagocytosis by white blood cells. These data are supported by the findings from several other laboratories in the field and previous results from our own research. This project is focused on testing and developing potential new therapeutic agents that will inhibit the bioactivity of the LOS by enzymatic removal of phosphate and acyl groups on the lipid and by inhibiting the genetic expression or competing for binding specifically to the active site of LOS biosynthetic enzymes. In addition, we will exten our observations regarding the inflammatory potential of the LOS and invasive ability of the Neisseria to interactions with an extensive set of relevant cell types that play a role in Neisseri infections. We will study models of infection in whole blood, in nasopharyngeal, cervical, and endothelial epithelial cells, monocytic cells, vascular endothelial cells, and brain cells, and analyze the bioactivity of LOS when inside of cells. We expect that the results will demonstrate the validity of new therapeutic targets, identify and develop potential new agents for the treatment of Neisseria infections, and increase our understanding of the complex relationship between these uniquely human pathogens and the innate immune system that enables the bacteria to cause disease.

Public Health Relevance

Our research focused on new therapeutics and innate immunity to Neisseria is highly relevant to the U.S. military and Veterans. Despite vaccination of all U.S. military personnel against N. meningitidis, disease incidence remains the same as that of the general population. From 2006-2010, 19% of the cases of meningococcal infection were fatal and 58% were vaccine failures. For gonorrhea, the U.S. rate remains high with ~820,000 new cases per year and antibiotic resistance is acutely problematic. Importantly, gonococcal infection is known to facilitate the transmission of HIV-1 infection. The high-risk environment of active military service is thought to increase risky behavior and, thus, the rate of sexually transmitted infections. Military and Veteran females, who incur a rate of gonorrhea that is higher than for civilian females and who suffer more frequent and serious complications than men, represent 15% of active duty military personnel and 2 million Veterans. Thus, our proposed research is relevant to the health and patient care missions of the VA.

Agency
National Institute of Health (NIH)
Institute
Veterans Affairs (VA)
Type
Non-HHS Research Projects (I01)
Project #
5I01BX000727-06
Application #
9262749
Study Section
Infectious Diseases B (INFB)
Project Start
2011-04-01
Project End
2020-06-30
Budget Start
2017-07-01
Budget End
2018-06-30
Support Year
6
Fiscal Year
2017
Total Cost
Indirect Cost
Name
Veterans Affairs Medical Center San Francisco
Department
Type
DUNS #
078763885
City
San Francisco
State
CA
Country
United States
Zip Code
94121
John, Constance M; Phillips, Nancy J; Din, Richard et al. (2016) Lipooligosaccharide Structures of Invasive and Carrier Isolates of Neisseria meningitidis Are Correlated with Pathogenicity and Carriage. J Biol Chem 291:3224-38
Gong, Zheng; Tang, M Matt; Wu, Xueliang et al. (2016) Arginine- and Polyamine-Induced Lactic Acid Resistance in Neisseria gonorrhoeae. PLoS One 11:e0147637
Liu, Mingfeng; John, Constance M; Jarvis, Gary A (2014) Induction of endotoxin tolerance by pathogenic Neisseria is correlated with the inflammatory potential of lipooligosaccharides and regulated by microRNA-146a. J Immunol 192:1768-77
Stephenson, Holly N; John, Constance M; Naz, Neveda et al. (2013) Campylobacter jejuni lipooligosaccharide sialylation, phosphorylation, and amide/ester linkage modifications fine-tune human Toll-like receptor 4 activation. J Biol Chem 288:19661-72
John, Constance M; Liu, Mingfeng; Phillips, Nancy J et al. (2012) Lack of lipid A pyrophosphorylation and functional lptA reduces inflammation by Neisseria commensals. Infect Immun 80:4014-26
Cheng, Hui; Yang, Zhijie; Estabrook, Michele M et al. (2011) Human lipooligosaccharide IGG that prevents endemic meningococcal disease recognizes an internal lacto-N-neotetraose structure. J Biol Chem 286:43622-33