Experience on Identification of Cross-Reactive Group Specificity Performed by Anti-human Globulin Panel Reactive Antibody Tests

Yong-Hak Sohn; Choong-Hwan Cha; Myeong Hee Kim; Sun-Young Ko; Heung-Bum Oh

doi:10.3343/kjlm.2008.28.5.362

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Sohn, Cha, Kim, Ko, and Oh: Experience on Identification of Cross-Reactive Group Specificity Performed by Anti-human Globulin Panel Reactive Antibody Tests

Original Article

Korean J Leg Med 2008;28(5):362-370.

Published online: 14 February 2008

DOI: https://doi.org/10.3343/kjlm.2008.28.5.362

Experience on Identification of Cross-Reactive Group Specificity Performed by Anti-human Globulin Panel Reactive Antibody Tests

Yong-Hak Sohn, M.D.¹, Choong-Hwan Cha, M.D.², Myeong Hee Kim, M.D.³, Sun-Young Ko, M.D.¹, Heung-Bum Oh, M.D.¹

¹Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea

²Department of Laboratory Medicine, University of Ulsan College of Medicine, Gangneung Asan Hospital, Gangneung, Korea

³Department of Laboratory Medicine, Kyung Hee University Medical Center, Seoul, Korea

Received 19 June 20085 August 2008 Accepted 19 August 2008

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

Panel reactive antibody (PRA) is to screen and identify HLA antibody. Majority of antibody specificities in high-PRA are directed against cross reactive group (CREG). Thus, this study was to know the advantage of identifying CREG specificity and whether antibody specificities are changed according to CREG classification.

Methods

HLA class I antibodies were identified from 159 sera from 108 patients in Asan Medical Center, who had shown more than 5% PRA by anti-human globulin (AHG)-complement-dependent cytotoxicity (CDC). Tail analysis-based computer program was developed to identify specificities, applying both Rodey (R-ABC) and Takemoto (T-ABC) classification. The results were also compared with those obtained when without CREG application (ABC).

Results

Among 151 cases in which HLA specificities was identified, the frequency of CREG specificity was 22.5% in R-ABC and 27.2% in T-ABC. Eleven cases showed CREG specificities only in one classification. However, the individual antigen specificities in one hand were all included in the CREG identified in the other hand. CREG specificities in samples with PRA >50% (60%) were more frequently identified than those in samples with PRA ≤50% (9%) (in R-ABC, P<0.0001). Without applying CREG to interpretation, specificity was not identified in 9 cases.

Conclusions

Application of CREG enhanced the rate of antibody identification. Antibody specificities of those cases where CREG specificities were different between Rodey and Takemoto classifications were almost the same when compared at the individual antigen level. Therefore, it was thought that it makes no difference to use any one of these two classifications in interpreting PRA.

Keywords: Panel reactive antibody, Tail analysis, Computer program, Human leukocyte antigen, Antibody specificity, Cross reactive group (CREG)

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Table 1.

HLA distribution of 11 PRA lots and allele frequencies in Korean population[22]

HLA-A	Allele frequency in Korean (%)	Lot (%)^∗	ID (%)^†	HLA-B	Allele frequency in Korean (%)	Lot (%)	ID (%)	HLA-C	Allele frequency in Korean (%)	Lot (%)	ID (%)
A1	1.46	4.68	7.45	B7	3.88	4.42	6.28	Cw1	19.58	18.18	4.76
A2	30.42	26.52	22.34	B8	0.32	1.39	7.11	Cw2	0.97	1.64	14.29
A3	1.13	2.53	1.06	B13	5.83	3.54	5.44	Cw3	24.11	24.75	9.52
A11	11.00	7.70	14.89	B14	1.13	2.27	0.42	Cw4	6.96	11.62	19.05
A24	22.81	22.85	25.53	B27	4.69	5.30	2.51	Cw5	0.81	1.14	28.57
A26	8.42	9.22	7.45	B35	6.95	6.44	4.18	Cw6	5.66	5.56	14.29
A29	0.32	0.88	0.00	B37	0.81	2.40	0.00	Cw7	11.17	14.77	4.76
A30	5.34	4.80	1.06	B38	1.13	1.39	2.09	Cw8	10.35	6.06	4.76
A31	4.37	4.30	8.51	B39	1.78	2.53	1.26	Cw12	2.92	2.91	-
A32	0.32	0.00	0.00	B44	8.41	7.83	7.53	Cw^∗14	15.04	10.73	-
A33	14.40	16.54	7.45	B46	6.15	5.05	4.60	Cw^∗15	2.42	0.88	-
A19	-	-	4.26	B47	0.16	0.00	0.00
				B48	2.75	1.52	4.18
				B50	0.16	0.88	3.77
				B51	12.79	8.71	8.37
				B52	2.43	2.15	2.09
				B54	5.02	6.44	2.09
				B55	2.59	3.54	1.67
				B56	0.48	0.00	0.00
				B57	0.32	0.00	0.00
				B58	5.83	6.82	2.93
				B59	1.46	2.53	4.18
				B60	3.56	5.31	4.18
				B61	7.77	4.67	1.26
				B62	9.55	9.97	2.93
				B67	0.81	3.03	3.35
				B71	1.78	1.14	3.35
				B75	1.29	0.25	0.42
				B5	-	-	5.86
				B15	-	-	0.00
				B16	-	-	0.00
				B17	-	-	0.00
				B22	-	-	1.26
				B40	-	-	6.69

^∗ , The proportion of the number of the corresponding antigen among a total number of antigens (36 × 2 × 11=792 antigens) in the individual locus of 11 PRA lots;

^† , The relative proportion of identified HLA specificity with the private or broad specificity in the individual locus (CREG specificities were not included).

Abbreviations: PRA, panel reactive antibody; CREG, cross reactive group; ID, identified

Table 2.

Frequency of identified antibody specificities to private, broad antigens and CREGs

Antigen	ABC^∗ (N=151)		R-ABC^† (N=151)		T-ABC^‡ (N=151)		Antigen	ABC^∗ (N=151)		R-ABC^† (N=151)		T-ABC^‡ (N=151)
Antigen	Antibody-identified person (N)	Frequency (%)	Antibody-identified person (N)	Frequency (%)	Antibody-identified person (N)	Frequency (%)	Antigen	Antibody-identified person (N)	Frequency (%)	Antibody-identified person (N)	Frequency (%)	Antibody-identified person (N)	Frequency (%)
A1	7	4.64	5	3.31	6	3.97	Cw3	2	1.32	0	0.00	0	0.00
A2	21	13.91	16	10.60	16	10.60	Cw4	4	2.65	5	3.31	5	3.31
A3	1	0.66	1	0.66	1	0.66	Cw5	6	3.97	5	3.31	5	3.31
A11	14	9.27	15	9.93	16	10.60	Cw6	3	1.99	0	0.00	0	0.00
A24	24	15.89	16	10.60	16	10.60	Cw7	1	0.66	2	1.32	2	1.32
A26	7	4.64	8	5.30	7	4.64	Cw8	1	0.66	1	0.66	1	0.66
A30	1	0.66	1	0.66	0	0.00	Unidentified	9	5.96	0	0.00	0	0.00
A31	8	5.30	8	5.30	8	5.30	Broad Ag	36	23.84	24	15.89	17	11.26
A33	7	4.64	5	3.31	4	2.65	A19	3	1.99	1	0.66	1	0.66
B7	15	9.93	11	7.28	11	7.28	B5	14	9.27	12	7.95	6	3.97
B8	17	11.26	13	8.61	15	9.93	B22	2	1.32	0	0.00	0	0.00
B13	13	8.61	10	6.62	11	7.28	B40	16	10.60	10	6.62	10	6.62
B14	1	0.66	1	0.66	1	0.66	A19+B22	1	0.66	1	0.66	0	0.00
B62	7	4.64	5	3.31	5	3.31	CREG	0	0.00	34	22.52	41	27.15
B75	1	0.66	1	0.66	1	0.66	A1C			1	0.66	0	0.00
B71	8	5.30	8	5.30	9	5.96	A2C			2	1.32	2	1.32
B27	6	3.97	2	1.32	2	1.32	B5C			6	3.97	8	5.30
B35	10	6.62	8	5.30	2	1.32	B7C			6	3.97	6	3.97
B37	0	0.00	1	0.66	1	0.66	B8C			1	0.66	0	0.00
B38	5	3.31	4	2.65	6	3.97	B12C			6	3.97	7	4.64
B39	3	1.99	3	1.99	3	1.99	Bw4C			9	5.96	10	6.62
B60	10	6.62	11	7.28	9	5.96	A2C+B5C			1	0.66	0	0.00
B61	3	1.99	3	1.99	3	1.99	A2C+Bw4C			1	0.66	1	0.66
B44	18	11.92	13	8.61	13	8.61	B5C+Bw6C			1	0.66	0	0.00
B46	11	7.28	13	8.61	13	8.61	B12C+Bw4C			0	0.00	1	0.00
B48	10	6.62	8	5.30	8	5.30	A10C					1	0.66
B50	9	5.96	8	5.30	8	5.30	(T-RBC only)
B51	20	13.25	19	12.58	19	12.58	B5C2					4	2.65
B52	5	3.31	5	3.31	5	3.31	(T-RBC only)
B54	5	3.31	6	3.97	7	4.64	A1C+A10C					1	0.66
B55	4	2.65	4	2.65	4	2.65	(T-RBC only)
B58	7	4.64	8	5.30	8	5.30	A2C+B5C2					1	0.66
B59	10	6.62	9	5.96	11	7.28	(T-RBC only)
B67	8	5.30	6	3.97	6	3.97	B5C2+Bw6C					1	0.66
Cw1	1	0.66	1	0.66	1	0.66	(T-RBC only)
Cw2	3	1.99	3	1.99	2	1.32

^∗ , HLA Class I specificities obtained without CREG application;

^† , Identified HLA Class I specificities according to applied Rodey classification;

^‡ , Identified HLA Class I antibody specificities according to applied Takemoto classification.

Abbreviation: CREG, cross reactive group.

Table 3.

Twelve cases showing different CREG specificity according to Rodey (R-ABC) and Takemoto (T-ABC) classification

Type	PRA%	ABC^∗						R-ABC^†						T-ABC^‡
R-ABC only	22	B27	B44	B38^§	B8	B59		B27	B44	B8C				B27	B44	B38	B8	B59
	36	B44	B13					B12C						B44	B13
	69	B5	A2	B35	B54			B5	A2	B35	B54			B5C	A2	B54
	50	B5	B35	B46	Cw5	B39	B71	B5	B35	B46	Cw5	B39	B71	B5C	B46	Cw5	B39	B71
	36	B5	B35	A11				B5	B35	A11				B5C	A11
	69^‖	A2	B5	B35				A2	B5	B35				A2	B5C
T-ABC only	72^‖	A2	B5	B35				A2	B5	B35				A2	B5C
	72^‖	A2	B5	B35				A2	B5	B35				A2	B5C
	39	A33	A26	B14				A33	A26	B14				A10C	B14
	81	A19	B22					A19	B22					A01C	A10C	B59	B71
	47	B44	A31	A30	Cw2	B8		B44	A31	A30	Cw2	B8		B12C	A31	B38	B8	A11
CREG difference	86	A19						A1C						Bw4C	B54

^∗ , HLA Class I specificities obtained without CREG application;

^† , Identified HLA Class I specificities according to applied Rodey classification;

^‡ , Identified HLA Class I antibody specificities according to applied Takemoto classification;

^§ , Private specificities which belong to the CREG specificities are marked in bold;

^‖ , Follow-up results from the same patient.

Abbreviations: See Table 1

Table 4.

The frequency of CREG specificity according to PRA%

Antibody specificity	R-ABC^∗,†		T-ABC^∗,‡
Antibody specificity	PRA>50% (N=40)	PRA≤50% (N=111)	PRA>50% (N=40)	PRA≤50% (N=111)
CREG ≥1^§	24 (60.0%)	10 (9.0%)	29 (72.5%)	12 (10.8%)
Private or broad only	16 (40.0%)	101 (91.0%)	11 (27.5%)	99 (89.2%)

^∗ , Both R-ABC and T-ABC have P<0.0001 by the Chi-square test;

^† , Identified HLA Class I antibody specificities according to applied Rodey classification;

^‡ , Identified HLA Class I antibody specificities according to applied Takemoto classification;

^§ , samples containing antibodies reactive to CREG specificities.

Abbreviations: See Table 1

Table 5.

HLA-C antibody specificities identified in 17 cases

No. Case	PRA%	Antibody specificities (R-ABC^∗)					Antibody specificities without inclusion of HLA-C in identifying antibody specificities using R-ABC
1	33	B40	B7	B48	Cw4		B40	B7	B48	B62
2	67	A2	Cw4	B5	B67	A11	A2	A11	B5	B67
3^†	22	B46	Cw5	A31			B46	A31	A3
4^†	58	A2	A24	Cw5			A2	A24	A3
5^†	8	B46	Cw5				B46	A3
6	75	Cw1	B5C	A26			B5C
7	75	Bw4C	Cw7	B48			Bw4C
8	50	B5	B35	B46	Cw5	B39	B5	B35	B46	B39	B71
9	8	B46	Cw2				B46	A3
10	33	Cw7	A31	B51	B35		B7
11	64	A2	Cw4	B51	A11		A2	B51	A11	B52
12	19	B7	Cw8	B60			B7	B60	B48	B54
13^†	47	B44	A31	A30	Cw2	B8	B44	A31	A30	B8	A3
14	36	B51	B58	Cw4			B51	B58	B14	A31
15	75	B7C	Cw4	B58			B7C
16	31	A33	Cw2				A33	B27
17	14	B13	Cw5	A31			B13	B62

^∗ , Identified HLA Class I antibody specificities according to applied Rodey classification;

^† , Follow-up results from the same patient;

^‡ , The results (B12C, A31, B38, B8, A11) in Takemoto CREG classification (T-ABC) didn't include HLA-C specific antibody.

Abbreviations: See Table 1

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