Corin L. Dorfmeier

Acidification of a solid sample to separate inorganic carbon (IC) from organic carbon (OC) is a widely encountered procedure in limnology and oceanography. Traditionally, OC was isolated to determine the weight‐% of OC (%OC), but it is becoming increasingly common to determine the isotopic signatures of the OC (δ 13 C OC and Δ 14 C OC ). This raises a need for a closer scrutiny of the currently used acidification methods. First, because IC and OC typically have distinct carbon‐isotopic signatures, δ 13 C OC and Δ 14 C OC values can be compromised if IC is not completely removed. Second, it is possible to isotopically fractionate a sample if a small portion of OC is lost during acidification, because OC itself is both chemically and isotopically heterogeneous. This study evaluated two acidification methods by HCl—in the vaporous (HCl vap ) and aqueous (HCl aq ) phases—to determine %OC, δ 13 C OC , and Δ 14 C OC in coastal sediments. Each method was assessed according to the criteria that it (1) has low blank levels, (2) is able to remove dolomite, (3) yields accurate %OC, and (4) yields accurate δ 13 C OC and Δ 14 C OC values. HCl vap fulfilled all criteria, given that the samples were not overexposed to acid. Overexposure led to underestimation of δ 13 C OC and Δ 14 C OC values. HCl aq gave similar results but was less reliable in that it consistently underestimated %OC and yielded inaccurate δ 13 C OC value for one test sample. It is recommended that an optimal acid exposure is carefully determined for each sample type to obtain most accurate δ 13 C OC and Δ 14 C OC values.

A major goal in rabies virus (RV) research is to develop a single-dose postexposure prophylaxis (PEP) that would simplify vaccination protocols, reduce costs associated with rabies prevention in humans, and save lives. Live replication-deficient RV-based vaccines are emerging as promising single-dose vaccines to replace currently licensed inactivated RV-based vaccines. Nonetheless, little is known about how effective B cells develop in response to live RV-based vaccination. Understanding this fundamental property of rabies immunology may help in developing a single-dose RV vaccine. Typically, vaccines induce B cells secreting high-affinity, class-switched antibodies during germinal center (GC) reactions; however, there is a lag time between vaccination and the generation of GC B cells. In this report, we show that RV-specific antibodies are detected in mice immunized with live but not inactivated RV-based vaccines before B cells displaying a GC B cell phenotype (B220(+)GL7(hi)CD95(hi)) are formed, indicating a potential role for T cell-independent and early extrafollicular T cell-dependent antibody responses in the protection against RV infection. Using two mouse models of CD4(+) T cell deficiency, we show that B cells secreting virus-neutralizing antibodies (VNAs) are induced via T cell-independent mechanisms within 4 days postimmunization with a replication-deficient RV-based vaccine. Importantly, mice that are completely devoid of T cells (B6.129P2-Tcrβ(tm1Mom) Tcrδ(tm1Mom)/J) show protection against pathogenic challenge shortly after immunization with a live replication-deficient RV-based vaccine. We show that vaccines that can exploit early pathways of B cell activation and development may hold the key for the development of a single-dose RV vaccine wherein the rapid induction of VNA is critical.

B cells secreting IgG antibodies, but not IgM, are thought to be solely responsible for vaccine-induced protection against rabies virus (RABV) infections in postexposure settings. In this report, we reinvestigated the potential for IgM to mediate protection in a mouse model of RABV vaccination. Immunocompetent mice immunized with an experimental live replication-deficient RABV-based vaccine produced virus neutralizing antibodies (VNAs) within 3 days of vaccination. However, mice unable to produce soluble IgM (sIgM(-/-)) did not produce VNAs until 7 days postimmunization. Furthermore, sIgM(-/-) mice were not protected against RABV infection when challenged 3 days postimmunization, while all wild-type mice survived challenge. Consistent with the lack of protection against pathogenic RABV challenge, approximately 50- to 100-fold higher viral loads of challenge virus were detected in the muscle, spinal cord, and brain of immunized sIgM(-/-) mice compared to control mice. In addition, IgG antibody titers in vaccinated wild-type and sIgM(-/-) mice were similar at all time points postimmunization, suggesting that protection against RABV challenge is due to the direct effects of IgM and not the influence of IgM on the development of effective IgG antibody titers. In all, early vaccine-induced IgM can limit dissemination of pathogenic RABV to the central nervous system and mediate protection against pathogenic RABV challenge. Considering the importance for the rapid induction of VNAs to protect against RABV infections in postexposure prophylaxis settings, these findings may help guide the development of a single-dose human rabies vaccine.