Kyushu University of Health and Welfare Repository

九州保健福祉大学生命医科学部生命医科学科・順正学園免疫学研究所順正学園免疫学研究所順正学園免疫学研究所departmental bulletin pape

池脇, 信直

イケワキ, ノブナオ

9000002154389

Ikewaki, Nobunao

1000090193226

90193226

村田, 興児

7925

日高, 直樹

7926

37J. of Kyushu Univ. of Health and Welfare. 23： 37〜41，2022
Serum titers of β-1,3-1,6 glucan antibodies were correlated with those of 
SARS-CoV-2 antibodies in healthy individuals
Ikewaki Nobunao 1）  Murata Koji 2）  Hidaka Naoki 2）
Abstract
　In this study, we examined if any correlations existed between the serum titers of β -1,3-1,6 glucan 
(BG) antibodies (BG-IgG, BG-IgM, and BG-IgA) and severe acute respiratory syndrome coronavirus-2 
(SARS-CoV-2) antibodies (SARS-CoV-2-IgG, SARS-CoV-2-IgM, and SARS-CoV-2-IgA) in 20 healthy 
individuals, using an enzyme-linked immunoassay (EIA). We found that BG-IgG and -IgM consistently and 
naturally existed in the sera of the study subjects; SARS-CoV-2-IgG and -IgM were also detected, albeit 
in low titers, in healthy individuals. However, no evidence of either BG-IgA or SARS-CoV-2-IgA was 
detected in the sera of the healthy individuals. The serum titers of BG-IgM were significantly correlated 
with those of SARS-CoV-2-IgM (r=0.636, P=0.015), whereas the serum BG-IgG titers were not 
significantly correlated with the serum SARS-CoV-2-IgG titers (r=0.214, P=0.365). Taken together, these 
findings suggest that antibodies to BG (mainly BG-IgM) may also play an important role in protecting 
against the development of coronavirus disease-19 (COVID-19), the disease caused by SARS-CoV-2 
infection.
Key words ：BG antibodies, COVID-19, SARS-CoV-2 antibodies, serum
Introduction
The corona virus disease-19 (COVID-19) is caused by 
the novel coronavirus, termed as severe acute 
respiratory syndrome coronavirus-2 (SARS-CoV-2). The 
clinical signs and symptoms of COVID-19 include low- 
to high-grade, unproductive cough, muscle pain, 
dyspnea, malaise, normal or decreased total leukocyte 
count, and evidence of pneumonia on the chest X-ray 1). 
However, some patients are asymptomatic, while 
others develop severe disease and die of the infection 2). 
The genetic , physiological and immunological 
mechanism(s) that could explain the wide spectrum of 
clinical severity of COVID-19 have yet to be elucidated. 
Recently, it was reported that acquired immunity to 
the common cold, also caused by a coronavirus, may 
afford protection against SARS-CoV-2 infection 3). Even 
persons who have never been infected with SARS-
CoV-2 have been demonstrated to show immunity to 
this virus, suggesting that antibodies raised against 
some environmental common bacteria, seasonal viruses 
and fungi may also react with SARS-CoV-2, through 
the phenomenon called cross-reactivity 3).
β-1,3-1,6 glucans (BG) are mainly found in the 
extracts of some species of mushrooms and in 
microbes such as the black yeast and candida, and 
possess some unique immunological activities. BG have 
been shown to exert cytotoxic activity against cancer 
cells through inducing the production of interleukin-2 
(IL-2), IL-6, IL-12, interferon-γ (IFN-γ), and tumor 
necrosis factor-α (TNF-α). These findings indicate that 
BG triggers immune responses, with the activation of 
lymphocytes, monocytes, macrophages, granulocytes, 
and natural killer (NK) cells 4-6).    
It has been reported that human sera consistently 
and naturally contain BG antibodies, suggesting that 
t h e BG a n t i b o d i e s may p l a y a n impo r t a n t 
immunological role 7). In particular, the BG antibodies 
may have the ability to protect against infectious 
diseases caused by various pathogens, including SARS-
1）Department of Medical Life Science, Kyushu University of Health and Welfare School of Medical Life Science, Institute of Immunology, 
Junsei Educational Institution, 1714-1 Yoshino-cho, Nobeoka-city, Miyazaki, 882-8508 Japan
2）Institute of Immunology, Junsei Educational Institution, 1714-1 Yoshino-cho, Nobeoka-city, Miyazaki 882-8508, Japan 
九州保健福祉大学研究紀要　23： 37〜41，2022
CoV-2. However, the associations and correlations of 
BG antibodies and SARS-CoV-2 antibodies are not yet 
unders tood . In th i s s tudy , we examined the 
correlations between the levels of BG antibodies and 
those of SARS-CoV-2 antibodies in the serum of 20 
healthy individuals.
Materials and Methods 
Reagents
SARS-CoV-2 (2019-nCoV) spike S1-his recombinant 
protein (Cat: 40591-V08H) was purchased from Cosmo 
Bio Co. (Tokyo, Japan). β -1,3-1,6 glucan (BG) was 
synthesized from Aureobasidium pullulans (A. 
pullulans ) using the latest biological culture and 
preparation techniques, and is currently available 
commercially as a health food supplement. In this 
study, the purified BG described above was kindly 
provided to us for this study by Sophy Co. (Kochi-city, 
Kochi). Horseradish peroxidase (HRPOD)-conjugated 
rabbit anti-human IgG, HRPOD-conjugated rabbit anti-
human IgM, and HRPOD-conjugated mouse anti-human 
IgA monoclonal antibody (mAb) were purchased from 
MBL Co. (Nagoya Japan). 
Ethics statement 
The study protocol was approved by the institutional 
review board (IRB) of Kyushu University of Health and 
Welfare; the IRB numbers were 20-020. Informed 
consent was obtained from all the blood sample donors 
prior to their participation in this study. 
Volunteers and collection of serum specimens
Blood samples of 20 normal healthy individuals (13 
males, age 38.2 ± 12.1 yr; 7 females, age 45.3 ± 6.2 yr) 
were collected at Suisonia Co. (Kitakyushu-city, 
Fukuoka, Japan). Serum was separated from the 
samples by the standard method. The samples were 
then centrifuged at 3,000 rpm for 30 min, and the 
supernatants were harvested. The serum samples 
were stored at -80℃ until the assays. 
 
Measurement of BG antibodies and SARS-
CoV-2 antibodies in the serum samples 
EIA plates (Sumitomo Co., Tokyo, Japan) were 
coated with 1µg/mL of purified BG or SARS-CoV-2 
spike protein (S1) in carbonate bicarbonate buffer (0.010 
M NaCO3, 0.035 M NaHCO3, pH9.6) and left to stand in 
for 24 h at 4℃ . The wells were then washed four times 
with phosphate-buffered saline (PBS) containing 0.05% 
Tween-20 (PBST) and blocked with PBST containing 2% 
BSA (BSA-PBST) for 60 min at room temperature. 
Thereafter, the wells were washed three times with 
PBST. Then 50µL of the serum samples (×1,000 dilution 
for IgG, ×500 dilution for IgM, and ×4 dilution for IgA 
detection of BG-antibodies, ×100 dilution for IgG and 
IgM, and ×4 dilution for IgA of SARS-CoV-2 antibodies) 
were then added to each well, followed by incubation of 
the plate for 60 min at room temperature with shaking. 
Then, the wells were washed five times with PBST, 
followed by the addition of HRPOD-conjugated rabbit 
anti-human IgG, HRPOD-conjugated mouse anti-human 
IgA mAb or HRPOD-conjugated rabbit anti-human 
IgM (each in ×5,000 dilution with 0.1% BSA-PBST; 
50µL) to each well, and the plates were incubated for 
60 min at room temperature with shaking. The wells 
were then washed 10 times with PBST, followed by 
the addition of a chromogenic substrate (TMB; Cosmo 
Bio Co.; 50 µL) to each well and incubation of the plate 
for 20 min at room temperature with gentle shaking. 
The reaction was stopped by the addition of 0.5M-HCl 
(50µL), and the optical density (O.D.) was read at 
450nm using a multichannel EIA-microplate reader 
(TOSHO Co.). The experiment was repeated five times. 
Statistical analysis 
Statistical analyses of the correlations of the serum 
levels of the BG-IgG, -IgM and -IgA with the serum 
levels of the SARS-CoV-2-IgG, -IgM and -IgA were 
performed by the Pearson correlation test. Differences 
at P<0.05 were considered as being indicative of 
statistical significance. 
38
Ikewaki Nobunao, Murata Koji, Hidaka Naoki ：Correlations between β -glucan antibody and SARS-CoV-2 antibody titers
Results and Discussion
In this study, we found that the BG-IgG and -IgM 
consistently and naturally existed in the sera of the 20 
healthy individuals, with the levels varying widely 
among individuals; the sera were also found to be 
weakly positive for SARS-CoV-2-IgG and -IgM (Fig.1). 
BG-IgA and SARS-CoV-2-IgA were almost never 
detected in the sera of the healthy individuals (Fig.1). 
As shown in Fig. 2, interestingly the serum levels of 
BG-IgM were significantly correlated with the serum 
levels of SARS-CoV-2-IgM (r=0.636, P=0.015). On the 
other hand, the serum levels of BG-IgG were not 
correlated with serum levels of SARS-CoV-2-IgG 
(r=0.214, P=0.365), suggesting that BG-IgM in the 
serum may play an important role in the protective 
immunobiological responses against SARS-CoV-2 
infection and development of COVID-19. 
In regard to the clinical manifestations of COVID-19, 
a wide spectrum of disease severity is observed, with 
some people remaining asymptomatic, but others 
developing severely illness or even dying from the 
d i s e a s e 1,2) .  Th e g en e t i c ,  p hy s i o l o g i c a l  a n d 
immunological mechanism(s) underlying such a wide 
spectrum of clinical severity of the disease are not yet 
understood at the present time. One possible important 
reason is the existence of cross-reactive antibodies in 
the body. Observations also suggest that several 
antibodies acquired by vaccinations, such as Bacille de 
Calmette et Guérin (BCG) vaccination, and influenza 
and measles-mumps-rubella (MMR) vaccinations may 
cross-react with SARS-CoV-2, accompanied by 
activation of CD4+ and CD8+ T-lymphocytes (cytotoxic 
T-lymphocytes; CTLs) that recognize these vaccine 
antigens 8,9). For example, it was reported that pre-
treatment of mice with attenuated Candida dubliniensis 
completely and effectively suppressed infections by the 
highly virulent Staphylococcus aureus and Candida 
albicans, suggesting that some cross-reactive antibodies 
or CD4+ and CD8+ T-lymphocytes (CTLs) induced by 
Candida dubliniensis  effectively protect against 
infections caused by more virulent organisms 10). 
In our study, the BG-IgM titers showed significant 
correlat ions with the SARS-CoV-2-IgM t iters , 
suggesting that BG-IgM cross-reacts with the SARS-
CoV-2 spike protein (S1) to neutralize it and prevent 
the onset of COVID-19. In fact, we found, for the first 
time in this study, that rabbit anti-BG-specific antibody 
cross-reacted with the SARS-CoV-2 spike protein (S1) 
in a dose-dependent manner (data not shown). 
BG is a component of mold/fungi, and we are 
routinely exposed to mold/fungi infection in the 
environment. In particular, Candida albicans is a 
popular mold that often thrives in living organisms, 
causing opportunistic infections in persons with 
impaired immunity. Serum samples from humans 
consistently and naturally contain BG antibodies 7), 
suggesting that BG antibodies7) may play an important 
immunological role in innate immunity and protect 
SARS-CoV-2 infection. It appears possible that healthy 
individuals with high BG-IgM titers are less susceptible 
to infection with SARS-CoV-2. In this study, since no 
correlation was observed between the titers of BG-IgG 
and SARS-CoV-2-IgG, the IgM subclass of antibodies 
may be predominantly and closely associated with 
protection against the development of COVID-19.
Although high-quality vaccine and antiviral agent are 
needed for the management/control of the COVID-19, 
BG-IgM in the serum as cross-reactive antibodies may 
protect against the development of COVID-19. Further 
studies are needed for detailed investigation of the 
immunological roles and mechanism underlying the 
effects of BG antibodies and SARS-CoV-2 antibodies at 
the cellular and molecular levels. 
Acknowledgements 
We wish to extend our gratitude to Hashimoto K of 
Suisonia Co. (Kitakyushu-city, Fukuoka, Japan) for 
preparing and providing us with serum samples of 
healthy individuals.
Disclosure 
None of the authors has any conflict of interest to 
disclose. 
39
九州保健福祉大学研究紀要　23： 37〜41，2022
References
1. Izda, V., Jeffries, M.A., Sawalhaet, A., H.: COVID-
19: A review of therapeutic strategies and vaccine 
candidates. Clin. Immunol. 222: 108634, 2021.
2. Majumder, J., Minko, T.: Recent developments on 
therapeutic and diagnostic approaches for COVID-
19. AAPS. J. 23: 14, 2021.
3. Lv, H., Wu, N.C., Tsang, O.,T. : Cross-reactive 
antibody response between SARS-CoV-2 and 
SARS-CoV infections. Cell Rep. 31: 107725, 2020.
4. Ikewak i , N . , Fu j i i , N . , Onaka , T . , e t . a l . : 
Immunological actions of Sophy beta-glucan (beta-
1,3-1,6 glucan), currently available commercially as 
a health food supplement.Microbiol. Immunol. 51: 
861-873, 2007.
5. Ikewaki, N., Iwasaki, M., Abraham, S.J.K.J : 
Biological response modifier glucan through 
ba l an c i ng o f b l o od g l u c o s e may have a 
prophylactic potential in COVID-19 patients. J. 
Diabetes Metab. Disord.19: 1-4, 2020.
6. Ikewaki, N., Iwasaki, M., Kurosawa, G., et. al.: beta-
glucans: wide-spectrum immune-balancing food-
supplement-based enteric (beta-WIFE) vaccine 
adjuvant approach to COVID-19. Hum. Vaccin. 
Immunother. 17: 2808-2813, 2021. 
 
7. Ikewaki, N., Sonoda, T., Miyazawa, Y., et. al.: 
Serum levels of β -1,3-1,6 glucan-specific antibodies 
and immune biomarkers in normal individuals. J. of 
Kyushu Univ. of Health and Welfare. 19: 87-94, 
2018.
8. Escobara, L.E., Molina-Cruzb, A., Barillas-Muryb, 
C . : BCG vacc ine pro tec t i on f rom severe 
coronavirus disease 2019 (COVID-19). Proc. Natl. 
Acad. Sci. U.S.A. 117: 17720-17726, 2020.
9. Katoh, S., Obayashi, T., Ganesh, J.S., et. al.: Cross-
protection induced by encephalitis vaccines against 
COVID-19 might be a reason for relatively lower 
mortality rate in some countries. Arch. Acad. 
Emerg. Med. 8: e54, 2020.
10. Fidel, P.L., Noverr, M.C.: Could an unrelated live 
attenuated vaccine serve as a preventive measure 
to dampen septic inflammation associated with 
COVID-19 infection? mBio.11: e00907-20, 2020.
Figure 1. Measurement of the BG-IgG, -IgM and -IgA 
and SARS-CoV-2-IgG, -IgM and -IgA titers in the sera 
of 20 healthy individuals. The measurements were 
performed using an EIA system (See Materials and 
Methods for details). Each measurement was repeated 
four times.
Figure 2. Analysis of the correlations of the serum 
levels of BG antibodies (IgG and IgM) and serum levels 
of SARS-CoV-2 antibodies (IgG and IgM) in the entire 
subject population (n=20). A: Correlation between the 
serum levels of BG-IgM and serum levels of SARS-
CoV-2-IgM (r=0.636, P=0.015) in the entire subject 
population. B: Correlations between the serum levels of 
BG- IgG and serum levels of SARS-CoV-2- IgG 
(r=0.214, P=0.365) in the entire subject population. 
40
Ikewaki Nobunao, Murata Koji, Hidaka Naoki ：Correlations between β -glucan antibody and SARS-CoV-2 antibody titers
健常人における血清中β-1,3-1,6 glucan抗体とSARS-CoV-2抗体の相関性
池脇 信直 1）　村田 興児 2）　日高 直樹 2）
1）九州保健福祉大学生命医科学部生命医科学科・順正学園免疫学研究所　〒882-8508　宮崎県延岡市吉野町1714-1
2）順正学園免疫学研究所　〒882-8508　宮崎県延岡市吉野町1714-1 
要旨
　健常人（20名）におけるβ -1,3-1,6 glucan（BG）抗体と SARS-CoV-2抗体の血清レベルを酵素抗体法（enzyme 
immunoassay：EIA）を用いて検討した。その結果、健常人血清中には BG-IgGおよび BG-IgMが恒常的に
存在することが分かった。また、力価は弱いが SARS-CoV-2-IgGおよび SARS-CoV-2-IgMも存在することが
分かった。しかしながら、BG-IgAおよび SARS-CoV-2-IgAはほとんど存在しなかった。次に、BG抗体と
SARS-CoV-2抗体との相関性を検討したところ、BG-IgMは SARS-CoV-2-IgMと統計学的に有意な相関を示
した（r=0.636,　P=0.015）。しかしながら、BG-IgGは SARS-CoV-2-IgGとは統計学的に有意な相関を示さなか
った（r=0.214,　P=0.365）。以上の結果は、BG-IgMは新型コロナウイルス感染症（COVID-19）の防御に何ら
かの関連性を有している可能性があることが示唆された。
キーワード ：BG 抗体、COVID-19、SARS-CoV-2 抗体、血清
41


Serum titers of β-1,3-1,6 glucan antibodies were correlated with those of SARS-CoV-2 antibodies in healthy individuals

山岸, 治男

7933

A Study of the course of formation of Anxiety about School Attendance from Psychosocial Point of View

2021年度
2022年３月
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ISSN 2434-2610


表紙

目　次
｢のべおか子どもセンター｣ 20年目の活動報告 …………………………………………… 1
　　　山本 順之、 松田 智香子、 井藤 英俊、 正野 知基、 天野 善昭、 二見　豊
若手保育士の仕事の困難さへの克服過程について ……………………………………… 21
　　　三宮 基裕、 黒木 香里


倉内 紀子（臨床心理学科）
佐々木さはら（スポーツ健康福祉学科） 　西田　美香（臨床福祉学科） 樋口　博之（作業療法学科）
川野　純一（視機能療法学科） 　砂子澤　裕（臨床工学科） 白﨑　哲哉（薬学科）
片本　　宏（動物生命薬科学科） 　野村　　創（生命医科学科） 吉武　重徳（臨床心理学科）
図 書・紀 要 委 員 会
図書・紀要委員会委員長
図 書 ・ 紀 要 委 員
九州保健福祉大学研究紀要
目　　　　次
第23号　令和４年３月
人文・社会科学
発達障害及び発達障害の疑いのある学生の言語聴覚士学外実習の様相と学習困難に関する調査
 　　　　　　　　　 ･･･････････････････････････････････････････････････････････ 松山光生 , 戸高翼 , 谷本愛裕美 , 中嶋将人  …  1
校則制定・改廃に果たす特別活動の意義と可能性に関する一試論 ･････････････････････････････････････････････登坂　学 … 11
陸上競技における種目間トランスファーを目的としたテストの開発 ･･････････････････････････････井藤英俊 , 山本順之 … 25
自然科学
薬学生を対象とする携帯型超音波診断装置を使用した膀胱内尿量測定の予備的試験とその意識調査
 　　　　　　　　　 ･････････････ 徳永仁 , 帳智香 , 河田実優 , 丸山凜七 , 園田純一郎 , 興梠靖幸 , 緒方賢次 , 髙村徳人 … 33
健常人における血清中β -1,3-1,6 glucan 抗体と SARS-CoV-2 抗体の相関性
 　　　　　　　　　 ･････････････････････････････････････････････････････････････････････池脇 信直 , 村田 興児 , 日高 直樹 … 37
新型コロナウィルス感染症拡大時（第 3 波）における宮崎県内在宅高齢者の生活状況調査
 　　　　　　　　　 ･････････････････ 原　修一 , 日田　剛 , 佐々木さはら , 井藤英俊 , 兒崎友美 , 川﨑順子 , 三浦宏子 … 43
ヒト血清アルブミンのサイトⅠおよびⅡに結合する薬物に対する長鎖脂肪酸と中鎖脂肪酸の影響
 　　　　　　　　　 ･････････････････････････････････････ 髙村徳人 , 緒方賢次 , 尾﨑峯生 , 石井紗綾 , 興梠靖幸 , 徳永仁 … 51
投稿規程
編集後記
Kurauchi Noriko
Sasaki Sahara Nishida Mika Higuchi Hiroyuki
Kawano Jun-ichi Isakozawa Yutaka Shirasaki Tetsuya
Katamoto Hiromu Nomura Hajime Yoshitake Shigenori
EDITORS
Editor in Chief
Editor
JOURNAL OF
KYUSHU UNIVERSITY OF HEALTH AND WELFARE
CONTENTS
No. 23 March 2022
Humanities and Social Science
Off-campus Speech-Language-Hearing Therapist training features and learning difficulties for students with confirmed 
or suspected developmental disabilities
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A Consideration on the Significance and Possibility of Special Activities for the Establishment, Revision and Abolition of 
School Regulations
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Natural Science
Preliminary study of intravesical urine volume measurement using a portable ultrasonic diagnostic device and a survey 
on the awareness about this device among pharmacy students
　　　　　　　　　　　 ･･････････････････････ Jin Tokunaga, Tomoka Cho, Miyu Kawata, Rina Maruyama, Junichiro Sonoda,  
　　　　　　　　　 Yasuyuki Kourogi, Kenji Ogata, and Norito Takamura … 33
Serum titers of β -1,3-1,6 glucan antibodies were correlated with those of SARS-CoV-2 antibodies in healthy individuals
　　　　　　　　　　　 ････････････････････････････････････････････････････････Ikewaki Nobunao, Murata Koji, Hidaka Naoki  … 37
Survey of the living conditions of community-dwelling older adults in Miyazaki Prefecture at the time of the spread of 
COVID-19 （third wave）
　　　　　　　　　　　 ･･････････････Shuichi HARA, Tsuyoshi HITA, Sahara SASAKI, Hidetoshi ITO, Yumi KOZAKI,
　　　　　　　　　 Yoshiko KAWASAKI, and Hiroko MIURA … 43
Effects of long- and medium-chain fatty acids on drugs that bind to sites I and II of human serum albumin
　　　　　　　　　　　 ･････Norito Takamura, Kenji Ogata, Mineo Ozaki, Saya Ishii, Yasuyuki Kourogi, Jin Tokunaga … 51
Contribution Rule
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九州保健福祉大学学芸員養成課程

7689

令和3 年度の博物館実習( 館園実習 ・学内実習・見学実習) について

九州保健福祉大学博士（社会福祉学）doctoral thesi

永井邦明

Japanese

介護を必要とする高齢者を対象とした生きがい就業支援の在り方 ～通所介護事業所をフィールドとした質的研究～

目　　次
原著論文
登校不安の形成過程に関する心理社会的一考察
................................................................................................山岸 治男　.1
博士論文の概要........................................................................................................................... . 9
修士論文の概要........................................................................................................................... .10
執筆規程・執筆要領...................................................................................................................... .12
編集後記.................................................................................................................................... .15
Contents
Original Articles
A Study of the course of formation of Anxiety about School Attendance from Psychosocial Point of View
.......................... Haruo　YAMAGISHI　　1
Abstracts in Doctoral Thesis......................................................................................................... .9
Abstracts in Master's Thesis........................................................................................................ .10
Instructions for How to write..................................................................................................... .12
Authors..................................................................................................................................... .15


山本, 順之

7724

松田, 智香子

マツダ, チカコ

1000020320484

20320484

井藤, 英俊

7723

正野, 知基

6306

ショウノ, トモキ

9000010565935

Shono, Tomoki

1000060241220

60241220

天野, 善昭

7941

二見, 豊

7763

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Serum titers of β-1,3-1,6 glucan antibodies were correlated with those of SARS-CoV-2 antibodies in healthy individuals

A Study of the course of formation of Anxiety about School Attendance from Psychosocial Point of View

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令和3 年度の博物館実習( 館園実習・学内実習・見学実習) について

介護を必要とする高齢者を対象とした生きがい就業支援の在り方～通所介護事業所をフィールドとした質的研究～

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