Significance of Autoantibody Detection in the Diagnosis of Facial

Dermatitis

Xiaofang Zhou

, Xiangfen Deng

, Hongzhi Gu

, Yan Xiao

, Ling Li

, Zhiqiang Song

Zhifang Zhai

and Huan Wang

h,*

Department of Dermatology, First Affiliated Hospital of Army Military Medical University, 400038, Chongqing, China

Keywords: Facial Dermatitis, Autoantibodies, Connective Tissue Disease, Autoimmune Disease.

Abstract: To explore the significance of autoantibody detection in the diagnosis and differential diagnosis of facial

dermatitis, the results of autoantibody detection in 57 patients with “facial dermatitis” who were first

diagnosed in our outpatient department from July 2019 to March 2021 were retrospectively analyzed in this

paper. It is found that autoantibody detection is of great significance in the diagnosis and differential diagnosis

of facial dermatitis in patients with facial dermatitis, which can avoid missed diagnosis and contribute to the

early diagnosis of connective tissue disease.

1 INTRODUCTION

Facial dermatitis is the most common disease in

dermatology. It has a variety of clinical

manifestations, a long course of disease and many

complicated pathogenic factors. These features have

a certain impact on the diagnosis and follow-up

treatment of patients. In severe cases, facial dermatitis

may affect life and work. Because the causes of facial

dermatitis are complex. They may related with

patients' living environment, such as ultraviolet light,

and their use of cosmetics, medicine, their anxiety

and depression. Besides, it may also occur on the

basis of other diseases, leading to misdiagnosis. For

example, it is reported that of systemic lupus

erythematosus (sle) may initially misdiagnosed as

facial dermatitis (Chen, Tu, Yan, et al. 2018, Li, Zhi

2010).

Therefore, facial dermatitis is clinically suspected

to be related to autoimmunity, and autoantibody

testing is conducted to further confirm or exclude

https://orcid.org/0000-0001-9459-6543

https://orcid.org/0000-0001-8221-8924

https://orcid.org/0000-0002-3141-894X

https://orcid.org/0000-0003-2457-6066

https://orcid.org/0000-0003-2174-1177

https://orcid.org/0000-0002-4895-5522

https://orcid.org/0000-0003-0636-4588

https://orcid.org/0000-0001-9459-6543

such etiology. This paper reviewed the results of

autoantibody detection in 57 patients with facial

dermatitis, and discussed the significance of

autoantibody in the diagnosis and differential

diagnosis of facial dermatitis, so as to avoid missed

diagnosis and contribute to the early diagnosis of

connective tissue disease.

2 BACKGROUND REVIEW

Facial dermatitis mainly refers to skin inflammation

that occurs on the face and is characterized by

pruritic, recurrent and chronic tendency, There are

various manifestations, including facial seborrheic

dermatitis, facial atopic dermatitis, facial contact

dermatitis, hormone-dependent dermatitis, psoriasis,

acne, rose acne, lichen planus, lupus erythematosus,

etc. The pathogenesis involves many factors such as

autoimmunity, allergy, infection and vascular

reactivity (Song, Hao 2017). Qu Yan et al. found that

496

Zhou, X., Deng, X., Gu, H., Xiao, Y., Li, L., Song, Z., Zhai, Z. and Wang, H.

Signiﬁcance of Autoantibody Detection in the Diagnosis of Facial Dermatitis.

DOI: 10.5220/0011372800003438

In Proceedings of the 1st International Conference on Health Big Data and Intelligent Healthcare (ICHIH 2022), pages 496-499

ISBN: 978-989-758-596-8

contact dermatitis and hormone-dependent dermatitis

are the most common types of facial dermatitis in the

clinical and pathogenic analysis of patients with facial

dermatitis. Skin patch test and prick test are effective

methods to find allergens. For contact facial

dermatitis, aromatic compounds are the main

allergens in the patch test. The highest positive rate of

prick test is dust mites and dust mites (Qu, Meng,

Yang, et al. 2016).

In addition to these pathogenic factors, facial

dermatitis may also be related to autoimmune itself.

So conducting autoantibody testing can further

confirm or rule out such causes to avoid missed

diagnosis and to help early diagnosis of connective

tissue diseases.

ANA is a general term for autoantibodies against

all antigen components in cells. As an important

biological marker of autoimmune diseases, It is

commonly seen in patients with systemic (non-organ-

specific) autoimmune diseases such as mixed

connective tissue disease (MCTD), systemic lupus

erythematosus (SLE), sjogren's syndrome (SS),

systemic sclerosis disease (SSc), polymyositis (PM)

(Chinese Journal of Laboratory Medicine 2018). High

titer of anti-U1-NRNP antibody has diagnostic

significance for MCTD. Anti-sm antibody is a highly

specific serological marker of SLE. Positive anti-SSA

antibody and/or anti-SSB antibody are serological

criteria for SS diagnosis. Anti-scl-70 antibody is a

serological marker in SSc classification criteria,

which is associated with poor prognosis, pulmonary

fibrosis and heart disease. Anti-centromeric protein

(CENP) antibody is a specific serologic marker of

localized SSc, suggesting a good prognosis. Ama-a2

with high titer is the characteristic autoantibody of

PBC, and the positive rate of anti-JO-1 antibody in

PM patients is about 25%-30% (Chinese Journal of

Rheumatology 2014). Li Dongjiao et al. also

proposed that the combined detection of ANA and

anti-ENA antibody has more clinical application

value in the early diagnosis of autoimmune diseases

than ANA primary screening alone (Li, Guan, Xie, et

al. 2020).

Therefore, in this study, ANA and specific

autoantibodies against target antigens were detected

in patients with clinically suspected autoimmune

diseases.

3 DATA AND METHODS

3.1 Clinical Data

From July 2019 to March 2021, in the dermatology

department of our hospital, a total of 57 patients were

initially diagnosed with facial dermatitis at their first

outpatient visit, including 3 males and 54 females.

Among them, 11 patients were less than 20 years old,

17 patients were 21 to 30 years old, 9 patients were

31 to 40 years old, 11 patients were 41 to 50 years old,

and 9 patients were more than 50 years old. Inclusion

criteria: ①The first diagnosis is supposed to be facial

dermatitis; ② The clinical data is complete; ③

Patients with incomplete clinical data are excluded.

3.2 Method

3.2.1 Indirect Immunofluorescence (IIF)

Detection of ANA in Serum Samples

ANA in serum samples was detected by antinuclear

antibody IgG detection kit (indirect

immunofluorescence method, Oumeng Medical

Laboratory Diagnosis Co., Ltd.).

Operation method: First, centrifuge venous blood

samples to be tested, dilute the serum to be tested into

1:100 when conducting qualitative testing and dilute

the sample by multiple with PBS Tween buffer to

determine the antibody titer; Second, add 25ul of the

diluted sample drop bu drop on the reaction zone of

the Hep-2 cell matrix slide; Third, place the slide

covered with bio-flake face down, cover it in the

groove of the sample plate, and incubate at room with

temperature of 18℃ to 25℃ for 30 minutes; Then,

use a beaker to hold PBS Tween buffer and use it to

rinse the slide like running water, and immediately

immerse it in a beaker containing PBS Tween buffer

for at least 5 minutes; Next, remove the slide from the

washing cup, wipe off the moisture on the back and

edge of the slide with absorbent paper, and add 20ul

goat anti-human IgG dropwise labeled FITC in the

reaction area of the clean sample plate, and incubate

at room with temperature of 18°C to 25°C for 30

minutes; Then use a beaker to hold PBS Tween buffer

and use it to rinse the slide like running water, and

immediately immerse it in a beaker containing PBS

Tween buffer for at least 5 minutes; And last, remove

the slide from the washing cup, wipe off the moisture

on the back and edge of the slide with absorbent

paper, and add 10ul mounting medium dropwise to

mount the slide.

Signiﬁcance of Autoantibody Detection in the Diagnosis of Facial Dermatitis

497

Reading the slide: Observe the stained image of

cellsunder a fluorescence microscope (OLYMPUS

BX53, 40×objective).

Judgment of results: qualitative and antibody titer

results are judged according to the specific

fluorescence model. Positive: the nucleus shows a

specific fluorescence model. Negative: the nucleus

does not show specific fluorescence.

The titer is defined as the highest dilution factor

that can be observed for a specific fluorescence

reaction compared with the reaction phase with the

negative serum diluted by the same multiple Ratio.

3.2.2 Western Blotting Method to Detect

Anti-ENA Antibody Profile in Serum

Samples

The anti-ENA antibody spectrum in serum samples

was detected using the anti-nuclear antibody

spectrum IgG detection kit (Western blot method,

Oumeng Medical Laboratory Diagnosis Co., Ltd.),

including uracil 1-low molecular weight

ribonucleoprotein (U1-nRNP), Sm, Sjogren’s

syndrome A and B (SS-A, SS-B), 11-16 peptide

complex antigen (PM-Scl), cytoplasmic histoacyl-

tRNA synthetase (Jo-1), Value-added cell nuclear

antigen (PCNA), histones, mitochondrial M2 (AMA-

M2), nucleosomes, centromere protein B (CENP B),

ribosomal p protein, dsDNA, DNA topoisomerase I

(Scl-70) .

Operation method: First, dilute the serum to be

tested with sample buffer in the ratio of 1:101, take

out the required membrane strip, put it into the

incubation tank with the numbered side of the

membrane strip facing up, add 1.5ml samples to the

incubation tank and aspirate the liquid in the

incubation tank after 5 minutes of incubation on a

rocking bed at room temperature; Then, add 1.5 ml of

the diluted serum sample to the incubation tank,

incubate for 30 minutes at room temperature (18°C-

25°C) on a rocking bed, aspirate the liquid in the tank,

and wash the membrane strips 3 times with 1.5ml of

washing buffer on the rocking bed, 5 minutes each

time, and add 1.5ml anti-human IgG labeled by the

diluted alkaline phosphatase to the incubation tank

and incubate at room temperature (18°C-25°C) on a

rocking bed for 30 minutes; And then aspirate the

liquid in the tank, wash the membrane strips 3 times

with 1.5ml of washing buffer on the rocking bed ,

each time for 5 minutes, and add 1.5ml substrate

solution to the incubation tank, incubate at room

temperature (18℃-25℃) for 10 minutes on a rocking

bed, then aspirate the liquid in the tank, and wash the

membrane strip in distilled water 3 times, 1minute for

each time, and last put the test film strip in the result

judgment template, and judge the result after air-

drying.

The result is judged according to the coloring

intensity of the antigen band and the coloring

intensity of the quality control band. Positive: the

coloring of the antigen band is medium to strong or

the same as the intensity of the quality control band.

Negative: a white band appears on the membrane

strip where the antigen is coated.

4 RESULTS

Among 57 patients, 14 patients were positive for

ANA, with a positive rate of 24.56%. Among them,

11 patients had nuclear granular karyotype and 7

patients, 3 patients and 1 patients with titers of 1:100,

1:320, and 1:1000, respectively. There are 2 cases

with cytoplasmic granular karyotype, with a titer of

1:320; 1 case with nuclear homogeneous karyotype,

with a titer of 1:100. Results of serum ANA and anti-

ENA antibodies in 57 patients with facial dermatitis

are shown in the table below. Among the 14 patients

with ANA positive, 8 were positive for anti-SS-A

antibody, 4 were positive for anti-PM-Scl antibody

and anti-histone antibody, 2 were each positive for

anti-U1-nRNP antibody and anti-SS-B, and anti-Sm

Antibody, anti-AMA-M2 antibody, anti-ribosomal

protein p antibody and anti-nucleosome antibody

were positive in 1 case each. The final diagnosis was

1 case of SLE, 2 cases of DLE, and 2 cases of

undifferentiated connective tissue disease.

Table 1: Results of serum ANA and anti-ENA antibodies in

57 patients with facial dermatitis.

Test items

ANA positive

（

n=14

）

ANA negative

（

n=43

）

Anti-SS-A

（

57.14

）

（

2.33

）

Anti-PM-Scl

（

28.57

）

（

16.28

）

Anti-histone

（

28.57

）

（

6.98

）

Anti-U1-nRNP

（

14.29

）

（

6.98

）

Anti-SS-B

（

14.29

）

Anti-Sm

（

7.14

）

Anti-AMA-M2

（

7.14

）

（

16.28

）

Anti-ribosomal

rotein p

（

7.14

）

（

2.33

）

Anti-nucleosome

（

7.14

）

Anti-PCNA 0

（

6.98

）

Anti-Jo-1 0

（

2.33

）

Anti-centromere 0

（

2.33

）

ICHIH 2022 - International Conference on Health Big Data and Intelligent Healthcare

498

Among the 43 patients with negative ANA, 7 were

positive for anti-PM-Scl antibody, 7 were positive for

anti-AMA-M2, 3 were respectively positive for anti-

U1-nRNP antibody, anti-PCNA antibody, and anti-

histone antibody, and anti-SS-A antibody, anti-Jo-1

antibody, anti-ribosomal protein p antibody and anti-

centromere antibody were positive in 1 case each.

Although some of the 43 ANA-negative patients are

positive for special autoantibodies, these patients

have not yet been diagnosed with autoimmune

diseases.

5 CONCLUSIONS

This article retrospectively analyzed the autoantibody

test results of 57 patients with facial dermatitis. Of the

57 patients with facial dermatitis, 14 were positive for

ANA, with a positive rate of 24.56%. Among the 14

patients with positive ANA, 1 case was diagnosed

with SLE, 2 cases was diagnosed with DLE and 2

cases of undifferentiated connective tissue disease;

Among the 43 cases of ANA negative, although some

specific autoantibodies are positive, these patients

have not yet been diagnosed with autoimmune

diseases.

In this study, 57 patients with facial dermatitis in

the outpatient department were tested by ANA and

ENA. 14 cases (24.56%) were positive for ANA.

Among them, 1 case was diagnosed as SLE, 2 cases

were DLE, 1 case was connective tissue disease, and

1 case of undifferentiated connective tissue disease.

This also shows that in this part of patients with facial

dermatitis, some of them are the early atypical

manifestations of autoimmune diseases. Although

some of the 43 ANA-negative patients are positive for

specific autoantibodies, these patients are currently

not diagnosed with the above-mentioned related

autoimmune diseases.

In this study, among the patients with positive

autoantibodies, although the results of ANA detected

by indirect immunofluorescence were not consistent

with the anti-ENA antibody spectrum detected by

western blotting, ANA positive patients may still

suffer from mixed connective tissue disease (Zhou,

Wang, Yang, et al, 2012), systemic lupus

erythematosus and other autoimmune diseases (Chen,

Chen, Zhang, et al, 2014). Zhang Heng et al. also

pointed out in their research reports that for patients

with suspected autoimmune diseases, the test results

of ANA and ENA may not be completely consistent,

and the simultaneous detection of ANA and ENA can

reduce the rate of missed diagnosis and play a guiding

role in the diagnosis of autoimmune diseases (Zhang,

Wu, Tian, et al, 2018).

In conclusion, some of the patients clinically

diagnosed with facial dermatitis can detect positive

autoantibodies, and some positive antibodies are

associated with autoimmune diseases to a certain

extent. The detection of autoantibodies in patients

with facial dermatitis is of great significance for the

diagnosis and differential diagnosis of facial

dermatitis, and is helpful for the early diagnosis of

autoimmune diseases and avoiding missed diagnosis.

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