Name: evaluation of host-pathogen responses and vaccine efficacy in mice
Uploaded: 2019-02-22T13:30:00
Description: Watch this Scientific Journal Video about Evaluation of Host-Pathogen Responses and Vaccine Efficacy in Mice at JoVE.com

This work was supported by 1R01AI125560-01 and start-up funds from The Ohio State University.

All experiments with live animals were conducted following a protocol approved by The Ohio State University IACUC in accordance with IACUC guidelines. C57BL/6 mice were used in all immunizations and infections. Both male and female mice are used in each group as per NIH guidelines. The number of animals per group was determined by power calculations based on the predicted differences in outcome among experimental groups. For example, 8 mice per group will yield 80% power at &#945; = 0.05 (2-sided) for a 2-sample t</e...

The comprehensive protocol described here to study vaccine-induced immunity to B. pertussis infection will also permit evaluation of host responses to a variety of other pathogens. The protocol discusses methods to deliver immunizations, determine vaccine efficacy following pathogen challenge, and parallel dissection of immune function. In adapting the protocol in order to study other pathogens, several parameters would need to be modified. These include, but are not limited to, the mode of animal anesthesia, va...

Vaccines represent one of the greatest public health achievements of the 20th century, yet we still do not fully understand the mechanisms by which successful vaccines stimulate protective immunity. The identification of molecular signatures (e.g., cell activation markers, expansion of cellular subtypes, and patterns of gene expression) induced after vaccination provides a plethora of information for predicting and generating an efficacious immune response. The complexity of host-pathogen responses cannot be adequately replicated using in vitro cell culture systems1. In vivo vaccine models are designed to concomitantly evaluate multiple immune cell types within the host. This provides an advantage when characterizing vaccine antigen processing and presentation, differential cytokine secretion, and expansion of immune cells. The protocol described here provides a detailed method to determine vaccine efficacy through evaluation of the systemic and local immune responses and quantification of pathogen burden in tissues of interest. The example provided here tests the efficacy of an experimental vaccine for the pathogen Bordetella pertussis (B. pertussis).
B. pertussis is a gram-negative bacterium that is the etiological agent of the respiratory disease whooping cough (pertussis)2,3. Close contact with infected individuals (symptomatic or asymptomatic) leads to transmission, colonization, and disease. Despite significant global vaccine coverage4, pertussis is considered a resurging disease in many nations around the world and is a major cause of preventable childhood deaths5,6,7,8. In 2015, B. pertussis and pertussis were included in the National Institute of Allergy and infectious Diseases (NIAID) emerging infectious pathogen/disease list, emphasizing the need for development of a better vaccine that confers long-lived protective immunity.
Currently, an active area of investigation to control pertussis resurgence is development of a next-generation acellular pertussis vaccine (aPV) with an optimal combination of novel adjuvants and antigens to mimic the immune response elicited by the whole-cell pertussis vaccine (wPV)9. Using the protocol described, we recently reported that the modification of a current FDA-approved aPV by the addition of a novel adjuvant, Bordetella colonization factor A (BcfA), resulted in more efficient reduction of B. pertussis bacterial load from mouse lungs10,11. This increased protection was accompanied by the skewing of an alum-induced Th1/Th2 immune response to the more protective Th1/Th17 immune profile10. This protocol is detailed and comprehensive, enabling the investigator to obtain maximal information through concurrent evaluation of host and immune responses to a variety of pathogens.
The protocol described here follows the representative vaccine schedule, shown in Figure 1, to ensure optimal host immune responses.

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			<td height="21" style="height:21px;width:333px;">2L induction chamber</td>
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			<td>any brand is appropriate</td>
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			<td style="width:175px;">Fisher</td>
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			<td style="width:175px;">Fisher</td>
			<td style="width:119px;">M2670-500G</td>
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			<td style="width:119px;">F-7002</td>
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			<td height="21" style="height:21px;width:333px;">Glutathione</td>
			<td style="width:175px;">Research Products Int</td>
			<td style="width:119px;">G22010-25</td>
			<td></td>
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			<td height="26" style="height:26px;width:333px;">Ascorbic acid</td>
			<td style="width:175px;">Research Products Int</td>
			<td style="width:119px;">A50040-500</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:333px;">RPMI 1640</td>
			<td style="width:175px;">ThermoFisher Scientific</td>
			<td>11875093</td>
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			<td height="21" style="height:21px;width:333px;">FBS</td>
			<td style="width:175px;">Sigma</td>
			<td style="width:119px;">F2442-500mL</td>
			<td>&#160;any US source, non-heat inactivated</td>
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			<td height="21" style="height:21px;width:333px;">gentamicin</td>
			<td style="width:175px;">ThermoFisher Scientific</td>
			<td>15710064</td>
			<td style="width:307px;"></td>
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			<td height="21" style="height:21px;width:333px;">B-mercaptoethanol</td>
			<td style="width:175px;">Fisher&#160;</td>
			<td style="width:119px;">BP176-100</td>
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			<td height="21" style="height:21px;width:333px;">15mL dounce tissue grinder</td>
			<td style="width:175px;">Wheaton</td>
			<td>357544</td>
			<td>any similar brand is appropriate</td>
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			<td style="width:175px;">Kontes/Sigma&#160;</td>
			<td style="width:119px;">Z359971-1EA</td>
			<td>any similar brand is appropriate</td>
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			<td style="width:119px;"></td>
			<td>any brand is appropriate</td>
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			<td height="21" style="height:21px;width:333px;">3mL syringes</td>
			<td style="width:175px;">BD bioscience</td>
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			<td height="21" style="height:21px;width:333px;">15mL conical tubes</td>
			<td style="width:175px;">Fisher&#160;</td>
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			<td style="width:119px;">C2170</td>
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			<td height="21" style="height:21px;width:333px;">60mm plates</td>
			<td style="width:175px;">ThermoFisher Scientific</td>
			<td style="width:119px;">130181</td>
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		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:333px;">48-well tissue culture plates</td>
			<td style="width:175px;">ThermoFisher Scientific</td>
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			<td height="42" style="height:42px;width:333px;">1mL insulin syringe 28G1/2</td>
			<td style="width:175px;">Fisher Scientific/Excel Int.</td>
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			<td></td>
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			<td height="21" style="height:21px;width:333px;">Mouse IFN-gamma ELISA Ready-SET-Go! Kit</td>
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			<td style="width:119px;">50-173-21</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:333px;">Mouse IL-17 ELISA Ready-SET-Go! Kit</td>
			<td style="width:175px;">Invitrogen / eBioscience</td>
			<td style="width:119px;">50-173-77</td>
			<td></td>
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		<tr height="22">
			<td height="22" style="height:22px;width:333px;">Mouse IL-5 ELISA Ready-SET-Go! Kit</td>
			<td style="width:175px;">Invitrogen / eBioscience</td>
			<td style="width:119px;">50-172-09</td>
			<td></td>
		</tr>
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evaluation of host-pathogen responses and vaccine efficacy in mice

Vaccines are a 20th century medical marvel. They have dramatically reduced the morbidity and mortality caused by infectious diseases and contributed to a striking increase in life expectancy around the globe. Nonetheless, determining vaccine efficacy remains a challenge. Emerging evidence suggests that the current acellular vaccine (aPV) for Bordetella pertussis (B. pertussis) induces suboptimal immunity. Therefore, a major challenge is designing a next-generation vaccine that induces protective immunity without the adverse side effects of a whole-cell vaccine (wPV). Here we describe a protocol that we used to test the efficacy of a promising, novel adjuvant that skews immune responses to a protective Th1/Th17 phenotype and promotes a better clearance of a B. pertussis challenge from the murine respiratory tract. This article describes the protocol for mouse immunization, bacterial inoculation, tissue harvesting, and analysis of immune responses. Using this method, within our model, we have successfully elucidated crucial mechanisms elicited by a promising, next-generation acellular pertussis vaccine. This method can be applied to any infectious disease model in order to determine vaccine efficacy.

The model described shows a method to evaluate vaccine efficiency and immune responses during host-pathogen interactions. Figure 1 depicts the representative vaccine schedule used to immunize and infect mice and harvest tissues for analysis. Figure 2 demonstrates the setup of the anesthesia system employed to induce mice, enabling investigators to deliver immunizations and bacterial inoculums. Figure 3</stron...

Watch this Scientific Journal Video about Evaluation of Host-Pathogen Responses and Vaccine Efficacy in Mice at JoVE.com

Evaluation of Host-Pathogen Responses and Vaccine Efficacy in Mice

Here we present an elegant protocol for in vivo evaluation of vaccine effectiveness and host immune responses. This protocol can be adapted for vaccine models that study viral, bacterial, or parasitic pathogens.

Vaccines are a 20th century medical marvel. They have dramatically reduced the morbidity and mortality caused by infectious diseases and contributed to a ...

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