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Establishing ELISA and ELISpot assays to analyze maternal immunity to Rotavirus infection on offspring

Sadik Mustafa, Asma (2023) MOBN03 20221
Degree Projects in Molecular Biology
Popular Abstract
Maternal boost early in life alter rotavirus specific immune response on infants

The first period of life for infants goes through by depending on their mother’s antibody transfer, primarily because of the infant’s immune system is still under development while being exposed to pathogens. Maternal antibody transfer is achieved through placenta during pregnancy and continues past birth through breastfeeding. During the nursing period, several vaccines such as rotavirus vaccine are provided to infants to protect them from potentially fatal infections. Rotavirus (RV) is a contagious virus that damages the villi of the small intestine, causing malabsorptive diarrhea and dehydration. Despite the availability of rotavirus vaccines and its... (More)
Maternal boost early in life alter rotavirus specific immune response on infants

The first period of life for infants goes through by depending on their mother’s antibody transfer, primarily because of the infant’s immune system is still under development while being exposed to pathogens. Maternal antibody transfer is achieved through placenta during pregnancy and continues past birth through breastfeeding. During the nursing period, several vaccines such as rotavirus vaccine are provided to infants to protect them from potentially fatal infections. Rotavirus (RV) is a contagious virus that damages the villi of the small intestine, causing malabsorptive diarrhea and dehydration. Despite the availability of rotavirus vaccines and its high efficacy in developed countries, the efficacy of the same vaccines is rather poor in low-income/developing countries where the disease burden is high. The low rotavirus vaccine efficacy has been correlated to the interference of maternally derived antibodies on the infant’s ability to develop own immune response against infection and vaccination. By establishing a pre-conception mouse model in our previous work in the lab, we aimed to determine how the maternal rotavirus specific antibodies, which was produced because of RV infection in mice prior to pregnancy, affects their offspring’s RV specific immune response to RV infection. The conceived offspring’s immune cell composition of the small intestine and mesenteric lymph node were analyzed by flowcytometry, and results showed an induced general B cell response in RV infected pups born to RV-immune dam, but the B cell response was not RV specific. The RV-infected pups were certainly protected from infection by the transferred maternal antibodies, as no diarrhea was not detected.

ELISA and ELISpot assays are highly sensitive techniques used in studying the B cell immune response, which are the cells that produce antibodies that protect against pathogen if its recognized. In this project, we sought to optimize these assays to study the maternally transferred antibodies to offspring by using same pre-conception mouse model. Through establishing cross-fostering experiment, that is RV-infected mothers nurse pups born from naïve mothers and vice versa, our results from ELISA show that two different types of antibodies (IgA & IgG) are transferred through breastmilk for protection against RV. Additionally, the RV specific immune response is induced late in pups raised by RV-dam. RV is composed of many epitopes that the immune system recognizes and produce antibodies against these specific epitopes. VP6-specific (dominant RV epitope) IgA antibodies were detected in RV infected mice even as late as 9 weeks post infection by optimizing ELISpot assay. To conclude, our results demonstrate that the RV maternal antibodies in breastmilk postpone the offspring’s specific immune response to infection, and that the response is toward the VP6 epitope.

Master’s Degree Project in General Molecular Biology 60 credits 2023
Department of Biology, Lund University

Advisor: Katharina Lahl & Konjit Getachew Muleta
Biomedical Centre, LU/ Department of Immunology (Less)
Please use this url to cite or link to this publication:
author
Sadik Mustafa, Asma
supervisor
organization
course
MOBN03 20221
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
9113608
date added to LUP
2023-04-18 16:20:17
date last changed
2023-04-18 16:20:17
@misc{9113608,
  author       = {{Sadik Mustafa, Asma}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Establishing ELISA and ELISpot assays to analyze maternal immunity to Rotavirus infection on offspring}},
  year         = {{2023}},
}