Chapter 29
Methods to Evaluate Immune Cell Recruitment and Cellular
Uptake and Distribution of Antigen Following Intramuscular
Administration of Vaccine to Mice
Gerard Agbayani, Felicity C. Stark, Bassel Akache,
and Michael J. McCluskie
Abstract
An effective vaccine depends on the stimulation of the immune system to generate effective antigen-specific
immune responses capable of neutralizing mediators of disease long after vaccination. However, the ability
of the vaccine to enhance immune parameters such as cell activation, cell recruitment and antigen uptake
shortly following administration contributes to the development of long-term responses directed toward
the antigen. Here, we describe a flow cytometry-based method to identify changes in immune cell profile
and assess cellular uptake and distribution of antigen following vaccination.
Key words Vaccines, Cell recruitment, Antigen uptake, Flow cytometry
1 Introduction
Intramuscular immunization is routinely used in the clinic due to its
relative ease and consistency during vaccine administration. Emulating the clinical vaccination route when evaluating vaccine candidates in preclinical animal models is important, as the route can
influence (1) the magnitude/quality of the immune response
against a disease, (2) the site of immune activation (i.e.,
mucosal vs. systemic), and (3) the risk for local or systemic adverse
effects. In addition to characterizing antigen-specific immune
responses (humoral and/or cellular), it is possible to evaluate
immune cell recruitment and cellular uptake of antigen at the
vaccination site and in distant tissues, particularly draining lymph
nodes (LNs), following intramuscular immunization.
Herein, we describe a mouse vaccination model using ovalbumin (OVA) as a model antigen and a novel class of archaeosome
adjuvant. However, the methods described are appropriate for
other antigens and adjuvants. OVA is a chicken egg-derived
Blaine A. Pfeifer and Andrew Hill (eds.), Vaccine Delivery Technology: Methods and Protocols, Methods in Molecular Biology,
vol. 2183, https://doi.org/10.1007/978-1-0716-0795-4_29, © Springer Science+Business Media, LLC, part of Springer Nature 2021
513
Methods to Evaluate Immune Cell Recruitment and Cellular
Uptake and Distribution of Antigen Following Intramuscular
Administration of Vaccine to Mice
Gerard Agbayani, Felicity C. Stark, Bassel Akache,
and Michael J. McCluskie
Abstract
An effective vaccine depends on the stimulation of the immune system to generate effective antigen-specific
immune responses capable of neutralizing mediators of disease long after vaccination. However, the ability
of the vaccine to enhance immune parameters such as cell activation, cell recruitment and antigen uptake
shortly following administration contributes to the development of long-term responses directed toward
the antigen. Here, we describe a flow cytometry-based method to identify changes in immune cell profile
and assess cellular uptake and distribution of antigen following vaccination.
Key words Vaccines, Cell recruitment, Antigen uptake, Flow cytometry
1 Introduction
Intramuscular immunization is routinely used in the clinic due to its
relative ease and consistency during vaccine administration. Emulating the clinical vaccination route when evaluating vaccine candidates in preclinical animal models is important, as the route can
influence (1) the magnitude/quality of the immune response
against a disease, (2) the site of immune activation (i.e.,
mucosal vs. systemic), and (3) the risk for local or systemic adverse
effects. In addition to characterizing antigen-specific immune
responses (humoral and/or cellular), it is possible to evaluate
immune cell recruitment and cellular uptake of antigen at the
vaccination site and in distant tissues, particularly draining lymph
nodes (LNs), following intramuscular immunization.
Herein, we describe a mouse vaccination model using ovalbumin (OVA) as a model antigen and a novel class of archaeosome
adjuvant. However, the methods described are appropriate for
other antigens and adjuvants. OVA is a chicken egg-derived
Blaine A. Pfeifer and Andrew Hill (eds.), Vaccine Delivery Technology: Methods and Protocols, Methods in Molecular Biology,
vol. 2183, https://doi.org/10.1007/978-1-0716-0795-4_29, © Springer Science+Business Media, LLC, part of Springer Nature 2021
513