1Department of Human Physiology, Faculty of Basic Medical
Sciences, School of Medicine, Federal University Otuoke, Yenagoa, Bayelsa
State, Nigeria
2Department of Human Kinetics, Health and Safety Studies,
Faculty of Natural and Applied Science, Ignatius Ajuru University of Education,
Port Harcourt, Rivers State, Nigeria.
Heterophile antibodies
are influenced by both genetic and environmental factors, just like other
antibodies. Heterophile antibodies, which are produced by some disorders
including infectious mononucleosis (IM), have been found to interfere with
the results of hormone assays. With respect to age, sex, species, and social
status, their serum titre levels vary. Ninety-six (96) children between the
ages of five and twelve who appeared to be in good health were chosen at
random. Out of the subjects, sixty (60) were males and thirty-six (36) were
women. Blood sample was collected by venipuncture of the antecubital vein of
the fore arm of children (5-12 years). By employing the double dilution
method, sheep red blood cells (SRBCs) were exposed to human sera in order to
cause an antigenic challenge. Heterophile antigen titer value of male and female school age children was
compared with one-way analysis of varience (ANOVA) and P value less than
0.05 was considered significant. The results shows that children of between
the ages of 5-12yrs possesses heterophile antigens on their red blood cells
(RBC) and that this heterophile antigen were observe more on male subjects
than female subjects. Hower, the difference between the mean titers of male and female subjects was not statistically
significance at P < 0.05. A statistically non-significant difference was
observed between heterophile antigen titre of male and female school age
children in Port Harcourt metropolis.
Accepted:20/05/2024
Published: 05/06/2024
*Corresponding Author
Azosibe,
Perowei
E-mail: peroweia@ otuoke.edu.ng
Phone: +2348106992668
Keywords: Heterophile
Antigen, Heterophile antibody, Sheep’s Red Blood Cell (SRBC)
INTRODUCTION
Common antigens (or antigens
of similar nature) that are possessed by a variety of phylogenetically
unrelated species are known as heterophile antigens. Heterophile
antigen induced the synthesis of heterophile antibodies. These antibodies can
react with certain antigens, which are quite different from, and
phylogenetically unrelated to the one which evoked antibody response.Heterophilic antibodies are endogenous, nonspecific
antibodies that bind to a variety of different antigens [8].
They
can be found in patients with autoimmune diseases, but can also be present in
other inflammatory diseases as well as in healthy individuals [1].
Chemically,
heterophile antigens are composed of lipoprotein-polysaccharide
complexes. There is a possibility of there being identical chemical
groupings in the structure of mucopolysaccharides and lipids[4,5,9].
Studies with heterophilic monoclonal antibodies
with human tissues shows that humans carry heterophile antigen and are located
in different tissues such as the stomach, pancreas, heart, kidney tissue etc.
Also, the distribution of binding antigens of each strain of antibodies was
different in the population.
This may be the
reason why some people are more susceptible to infection with the same
pathogenic microorganism [2].
Furthermore,
several evidence supports the notion that sex chromosomes and gonadal hormones
modulate the number and functions of immune cells. There are well characterized
sex differences in the innate and adaptive immune response; there is strong
evidence that type I and type II interferon signaling and humoral responses are
greater in females than in males across diverse species. Sex differences in
both innate and adaptive immunity contribute to the increased prevalence of
autoimmunity in females and increase the propensity of females to reject their
organs post-transplantation. Sex differences research is uncovering novel
therapeutic pathways that could be targeted to improve disease outcomes in all sexes [2]. Thus, the need for this study.
MATERIALS AND METHOD
Research Population
and Study Location
Ninety-six (96)
apparently healthy children between the ages of 5-12 years were randomly
selected. Thirty-six (36/37.5%) of the subjects were female and sixty (60/62.5%)
males. The subjects were randomly drawn from government secondary schools, Port
Harcourt, Rivers State. The study was carried out within a period of five weeks
at the Department of Human Physiology, Faculty of Basic Medical Sciences,
College of Health Science, University of Port Harcourt.
Ethical Adherence
The University of Port Harcourt Research
Ethics Committee granted ethical permission.
The subjects, parents, and guardians were
given consent letters in which the goals, procedures, and advantages of the
study were spelled out in detail. The consent of parents or guardians was
obtained for the collection of blood samples from their children or wards, and
their individuals' identities were kept private [6].
Preparation of Sheep’s Corpuscle’s
Preservative (Alsever’s Solution)
The salts were measured out using a chemical
weighing balance (Mettlo Toledo-PL 2003) and the solution was prepared by mixing
20.5g dextrose, 8g sodium citrate, 0.552g citric acid, and 4.2g sodium chloride
in a little amount of distilled water and made up to 1000mL. Then the pH of the
solution was adjusted to 6.1. The solution was then divided into aliquots of
25mL, 25mL, and 150 mL and shared into 3 conical flasks respectively. The solutions
were autoclaved for 20 min at a pressure of 517mmHg for sterilization and
stored at a temperature of 4 ºC.
Collection of Sheep Red Blood Cells
The properly feed sheep was held very firm in
a place and furs at the tibial veinregion was shaved, ensuring that the tibial veins are visible and the
area swabbed with cotton wool soaked in methylated spirit .A 25 G vertinary
needle is inserted into the tibial vein and the (25ml) of venous blood from the
sheep is drawn into the aliquot bottle containing alservers solution making the
solution up to (50ml).The conical flask containing the mixture (alservers
solution)was then covered with a cling
film and carefully shake and stored in a refrigerator to age (3 days).
Experimental Design
Blood sample was collected by venipuncture of
the antecubital vein of the fore arm of children (5-12 years). Antigenic challenge
was induced by exposing the human sera to sheep red blood cells (SRBCs) using
the double dilution technique.
Assay of Heterophile
Antibodies
The anticoagulant known as Alsever's solution
was used to preserve SRBCs. To separate the SRBCs from the anticoagulant, this
was spun for five minutes at 3000 rpm. To obtain a clean supernatant, SRBCs
were spun three times after being cleaned in regular saline. Two drops of
regular saline were added to each well in the hemagglutination tray (including
the control wells) using a smooth-edged Pasteur pipette. Two drops of the test
sample (human serum) were introduced to the ninth well (positive control), and
two more drops were added to the first well and thoroughly mixed. Care was
taken in order to reduce bubbles. Two drops of the mixture were transferred from
the first well to the second, then from the second well to the third, and so
on, up to the eighth well (1/256), in order to make dilutions. Following
thorough mixing in the eighth and ninth wells, two drops were pipetted and
discarded to provide an equivalent volume of diluted serum. This process was
carried out again for every tray that was available. Every well, including the
control wells, had two drops of 1% SRBCs added to it, which included doubling
dilutions of the test samples. The samples were kept overnight at 4 ºC after
being blended to homogeneity by rocking and covered with cling film to avoid
evaporation. The titre values for the antigen-antibody reaction were displayed
in the agglutination-filled wells. Results were taken by viewing the wells with
mat appearance (just like the positive control) as positive (+) while those
with button appearance (just like the negative control) as negative. The
positive reading with the highest dilution of each row of wells was taken as
the titre value for that serum samples [4].
RESULTS
The results shows that children of between
the ages of 5-12yrs possesses heterophile antigens on the red blood cells (RBC)
and that this heterophile were observe more on male subjects than female
subjects. Hower, the difference between the mean titers of male and female
subjects was not statistically significance at P < 0.005 as shown in Table 1.
Table
1. Heterophile antigen titres and age of male and female apparently healthy
school age children
Statistics
Parameters
Sample size (N)
Mean ± SD of titre value
Mean ± SD of subjects age
Female (5-12yrs)
36 (37.5%)
30.83±49.279
7.42±2.383
Male (5-12yrs)
60 (62,5%)
53.20±89.296
8.47±2.288
DISCUSSION
The innate and adaptive immune systems
exhibit well-documented sex-based variations; type I and type II interferon
signaling as well as humoral responses are clearly higher in females than in
males across a wide range of animals. Innate and adaptive immunity differs
between the sexes, this is clearly in the likelihood that autoimmunity will
develop more often in females than males and transplanted organ rejection is
observed more in females. Research on gender differences is identifying new
therapeutic targets that may be used to enhance disease outcomes in both sexes [2,7].
There are variations
among species in the distribution of heterophilic antigens. Using matching
tissue microarrays from other animal species, immunohistochemical staining of fourteen
(14) heterophilic antibodies that reacted with antigens in human stomach,
pancreas, and kidney tissues showed that these antibodies likewise exhibited
distinct response features. These findings revealed that distinct individuals
had distinct distributions of heterophilic antigens, which could account for
why some people who are infected with the same harmful bacteria become ill
while others do not [2,3]. Also, research reports identified different
animals, organ systems, ages, environmental dispositions, and sexes to have
distinct immune responses. Different species'
neuro-immuno-endocrine pathways are influenced by the sex-determining
factor of their chromosomes [2,3]. Although the differences between
the heterophile antigen titres of male and female school-age children were not
statistically significant at P<0.05, this could be the cause of the
disparities. A larger sample size study will better address this problem.
CONCLUSION
A statistically non-significant difference
was observed between heterophile antigen titre of male and female school age
children in Port Harcourt metropolis.
2.Dunn, S.E., Perry, W.A. & Klein, S.L.
Mechanisms and consequences of sex differences in immune responses. Nat
Rev Nephrol 20, 37–55
3.Sun, L., Wang, N., Feng, Y., Huo, X., Feng,
Q., Zhao, X., ... & Hu, J. (2023). The distribution of heterophilic
antigens and their relationship with autoimmune diseases. Frontiers in
Immunology, 14, 1275658
4.Adeniyi-Jones C. (1976). Agglutination of
tanned sheep red blood cell of serum from Nigeria adults and children. Lancet,
1: 188-190.
6.Nuffield Council on Bioethics, 2011, The
Ethics of Research Related to Healthcare in Developing Countries. London:
Nuffield Council on Bioethics.
7.Udom, g. J., onyeukwu,
n. J., udobang, j. A., obot,
d. N., & yemitan, o. K. (2020). Immunotoxicological response of school age children to
environmental pollution and heterophile antigens in rivers state, southern nigeria. Journal of global biosciences, 9(6),
7586-7596.
8.Votava, M., Bartosová, D., Krchnáková, A., Crhová, K., & Kubinová, L.
(1996). Diagnostic importance of heterophile
antibodies and immunoglobulins IgA, IgE, IgM and low-avidity IgG
against Epstein-Barr virus capsid antigen in children. Actavirologica, 40(2), 99–101.
Cite this Article:Azosibe, P; Onesimus, ML; Azibaobom, K;
Ikete, PW; Adienbo, EN (2024). Sex Induced Disparity in Serum
Level of Heterophile Antibody in Apparently Healthy School-Age Children in
Port Harcourt, Rivers State, Nigeria. Greener
Journal of Medical Sciences, 14(2): 56-59.
