INTRODUCTION:

In 1950, this form of hepatitis was seen in a woman named Jan Waldenstrom. The presence of antinuclear antibodies gave the name lupoid hepatitis to this disease. This gave an idea about the immunological relationship of this disease. The name AIH was given by Mackay during 1965.It was recognized in an international meeting [1].

This disease was seen more in women than in men. The ratio of the number of infected women and men is 4: 1. The disease was seen mostly among teenage people and people who were at their menopause. But the fact is that the disease is seen in all age groups irrespective of the gender difference [2].

In the case of AIH, the parenchyma layer of the liver is destroyed and fibrosis develops. This makes the disease a serious problem. According to the data provided by WHO, one to two lakhs of Americans is affected by this disease. Fifteen among one lakh people are affected by this disease worldwide [3]. The reason for the affection of this disease is still not clear. However, the disease severely affects hepatocyte cells. This disease cause inflammation in the liver. This disease was initially identified in a woman named Waldenstorm during 1950. This condition leads her to liver cirrhosis. During the biopsy of the liver, morphological changes were observed. ANA was also identified in the serum [4]. The condition was seen more among women irrespective of their age. From blood tests elevation in the levels of various transaminase enzymes was observed. An increased level of immunoglobulin G antibodies was seen among these patients. Elevated levels of other antibodies like ASMA and LKM were also identified. Substances like toxins and drugs should not be there in the body of patients suffering from a severe form of hepatitis. The relapse rate of AIH affected men was high [5]. The patients suffering from this disease can be lead to cirrhosis. A strong relationship between HLA DR molecules was identified and is related to the increased number of AIH affected patients in Europe and North America. This relationship between HLA DR particles is helpful in the detection of HLA by the revised diagnostic scoring system implemented by IAIHG [7].

PATIENTS AND METHODS:

Two groups of people were used for the study of AIH disease. The first group consisted of sixty Iraqi people. Nineteen males and forty-one females were there in this group. The average age of the individuals belonging to this group was 40.6.All of them were confirmed to have AIH disease after conducting several tests [8]. There were fifty healthy individuals in the control group. There were fifteen males and thirty-five females in this group. All of them were Iraqi people having ages ranging from 10 to 67.They were subjected to several tests and was confirmed that they weren’t affected by AIH disease. Individuals from both the groups were subjected to ELISA tests and a German-made tool kit was used [9]. Using the primer supplied by the Korean Bioneer company, investigations for the appearance of HLA DR3 was conducted [10].

Table (1): Primer sequence with orientation and the PCR product size

HLA allele specific primer	Sequence		PCR Size
HLA-DR3	F	TTGTTGGGGTTCAAGTGG	80bp
	R	AAGCCACAADCCTGTTTTCC

Using SPSS software associated with Microsoft Excel, statistical analysis of the data was conducted. By using the odds ratio, the relationship between disease status and the appearance of the HLA genotype was studied [11].

RESULTS AND DISCUSSION:

From the data available from table 2, the results of the current study revealed that HLA is a condition characterized by the inflammation of the liver. This can affect different people irrespective of their age and sex. The study was conducted using two groups. The patients' group contained HLA affected patients having age between seven and sixty-nine. Their mean age was 40.6.The second group contained healthy individuals having their ages between ten and sixty-seven. The average age of people in the second group was 36.7. These data are similar to those reported by Mauss. Younger people are affected by AIH more [7]. This is mainly due to malnutrition and other environmental factors. It’s also influenced by the fact that the lifetime of Iraqi people is lower than Europeans. From table two, it is clear that females are affected by AIH more [12]. These results are similar to those of Baranov and Sabari. Variations seen in age is due to the influence of race and genetic factors. The hormonal variations in females activate Th2 and cause the production of autoantibodies. Because of this, AIH is seen more in females than in males [13].

Table (2): Distribution of studied groups by age and gender

Parameters	Healthy controls		Autoimmune		P (t-test)
Age (years)					0.17[NS]
Range	(10 to 67)		(7 to 9=69)
Mean	36.7		40.6
SD	15.0		14.9
SE	2.1		1.9
N₀	50		60
Gender	N	%	N	%	P(Chisquare) = 1[NS]
Female	34	68	41	68.3
Male	16	32	19	31.7
Total	50	100	60	100

These results are comparable with Goldsby et al. Anti SLA antibody details are provided in table three. We can see a significant difference in the level of these autoantibodies between the patients' group and the control group [14]. The level of such antibodies is more in AIH disease affected patients than in healthy people. 0.2 to 174.3 U/ml anti-SLA was seen in AIH affected group. 0.8 to 17.8 U/ml anti-SLA was seen in healthy individuals.

Table (3): Different (anti-SLA) level parameters among study groups.

Anti-Soluble Antibody (SLAU/ML)	Study group		P value
	Healthy controls	Autoimmune hepatitis	0.005
Range	(0.2 to 44.5	(0.2 to 174.3)
Median	4.6	12.4
Inter-quartile range	(2.8 to 13.1)	(5.5 to 22.3)
N0	50	60
Mean rank	23.7	35.4

All this information is displayed in the first figure. ROC curves were used for analyzing the diagnostic value of autoantibodies.

Table (4): Different (anti – AMA) level parameters among study groups.

Anti-Soluble Antibody (SLAU/ML)	Study group		P value
	Healthy controls	Autoimmune hepatitis	0.33[NS]
Range	(0.1 to 41)	(0.1 to 90.5_
Median	3.8	4.7
Interquartile range	(1.8 to 7.2	(1.05 to 14.8)
N	50	60

These studies showed an ROC value of 0.655 for this kind of autoantibody [15]. This is used as a test for the detection of AIH infection. These data are available in the fifth table.

Table (5): ROC area when used as test to predict a diagnosis autoimmune hepatitis differentiating them from healthy controls.

Auto-antibody (U/ml)	ROC AREA	P
SLA	0.655	0.005
AMA	0.554	0.33[NS]

The levels of AMA antibody among autoimmune hepatitis patients was not significant. Its concentration among healthy people was 3.8 U/ml. These data are available in table 4.These results are comparable to the results depicted by Mohamed. The differentiating value of these two autoantibodies has been calculated using ROC curves [16]. In the differentiation of autoimmune hepatitis patients from healthy individuals, the ROC area values are provided in table 5.

There are specific HLA genes that are related to the development of AIH disease [22]. The distribution of these alleles among the patients' group and the control group is given in the sixth table. The studies conducted about the distribution of the HLA DR3 genes revealed a relationship between the specific alleles and development of AIH conditions [17]. Considering the HLA DR3 genotype, 58.3% of patients showed a positive relationship with OR value 7.35.These values are obtained in comparing AIH patients with healthy individuals [18]. This information is not agreeing with ngu. After conducting the tests, it is clear that the presence of the HLA genotype increases the chance of having the disease [24]. The accuracy of this prediction was 79.1%. When the HLA DR3 genotype was seen at a younger age, those patients didn’t respond well to therapy [19]. This is associated with liver failure and required liver transplantation. This information is present in table 7.

Table (7): Multiple logistic regressions with the risk of being a case (autoimmune hepatitis) as the dependent (outcome) variable is HLA-DR3

HLA-gene	Partial OR	95% confidence interval	P
Positive DR3	14.7	(4.4 to 48.8)	<0.001

Overall predictive accuracy = 79.1%

P (Model) <0.001

The positive and negative distribution of serum in people with HLA genotype is included in table 8.Positive distribution of AMA serum was seen in patients with HLA DR3 genotype in a significantly higher manner than those who had negative AMA [20].

Table (8): Association of studying assay level with HLA-DR3

HLA-DR3
	Negative N(25)	Positive N(35)	P
AMA	0.006
RANGE	(0.1 to 60.3)	(0.1 to 90.5)
Median	2.1	11.4
Interquartile range	(0.8 to 10.3)	(2.7 to 23
SLA	0.001
RANGE	(0.2 to 44.5)	(0.2 to 174.3
Median	4.6	12.4
Interquartile range	(2.8 to 13.1)	(5.5 to 22.3)

The median of positive patients was 11.4 and for negative patients 2.1. In the case of SLA antibodies, the median recorded for positive patients was 12.4 and for negative patients, it was 4.6,with a p-value of 0.001.

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Received on 11.01.2020 Modified on 23.01.2020

Research J. Pharm. and Tech 2020; 13(2):819-822.

DOI: 10.5958/0974-360X.2020.00154.7

	Healthy controls (N= 50)		AIH patients (N=60)
HLA-DR3	N	%	N	%	OR	95%CI OR	P	EF
Positive	8	16	35	58.3	7.35	(2.95 -18.3)	<0.001	0.504
Negative	42	84.0	25	41.7