In Vivo Adaptation and Persistence of Neisseria meningitidis within the Nasopharyngeal Mucosa

Abstract

Neisseria meningitidis ( Nme) asymptomatically colonizes the human nasopharynx, yet can initiate rapidly-progressing sepsis and meningitis in rare instances. Understanding the meningococcal lifestyle within the nasopharyngeal mucosa, a phase of infection that is prerequisite for disease, has been hampered by the lack of animal models. Herein, we compare mice expressing the four different human carcinoembryonic antigen-related cell adhesion molecules (CEACAMs) that can bind the neisserial Opa protein adhesins, and find that expression of human CEACAM1 is necessary and sufficient to establish intranasal colonization. During infection, in vivo selection for phase variants expressing CEACAM1-specific Opa proteins occurs, allowing mucosal attachment and entry into the subepithelial space. Consistent with an essential role for Opa proteins in this process, Opa-deficient meningococci were unable to colonize the CEACAM1-humanized mice. While simple Opa-mediated attachment triggered an innate response regardless of meningococcal viability within the inoculum, persistence of viable Opa-expressing bacteria within the CEACAM1-humanized mice was required for a protective memory response to be achieved. Parenteral immunization with a capsule-based conjugate vaccine led to the accumulation of protective levels of Nme-specific IgG within the nasal mucus, yet the sterilizing immunity afforded by natural colonization was instead conferred by Nme-specific IgA without detectable IgG. Considered together, this study establishes that the availability of CEACAM1 helps define the exquisite host specificity of this human-restricted pathogen, displays a striking example of in vivo selection for the expression of desirable Opa variants, and provides a novel model in which to consider meningococcal infection and immunity within the nasopharyngeal mucosa.

Author Summary

Neisseria meningitidis ( Nme), a common cause of bacterial meningitis, are carried asymptomatically in the nasopharynx by a substantial proportion of healthy individuals. Their strict adaptation to the human as host has so far impeded the development of animal models to study the meningococcal lifestyle in vivo. While several human CEACAMs are recognized by the neisserial Opa protein adhesins, we show here that the expression of human CEACAM1 in transgenic mice is necessary and sufficient to allow nasal colonization by Nme. The dependence on human CEACAM1 is attributable to the Opa proteins, since intranasal infection with Opa-negative colonies of Nme selects for bacteria expressing Opa proteins, and genetically Opa-deficient meningococci are unable to colonize these animals. We use this new mouse model to examine how innate immune factors such as neutrophils and complement limit colonization. Furthermore, we compare how adaptive responses elicited by colonization and those generated by parenteral vaccination differentially confer sterilizing immunity. Together, this work provides the first evidence of the critical nature of Opa-CEACAM1 binding in vivo, demonstrates that this is a major determinant of the host restriction by Nme, and reveals a clear disparity between immune correlates of sterilizing immunity conferred by natural colonization versus parenteral immunization.