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Monoclonal Rh antibodies
Revue Franqaise de Transfusion et Immuno-h6matologie Tome XXXI. - N°2. - 1988
167
Monoclonal Rh antibodies by P. Tippett and C. Lomas MRC Blood Group Unit, Wolfson House, 4 Stephenson Way, London NW1 2HE, U.K.
The Rh antibodies were studied by m a n u a l serological techniques (see Daniels' report on Kell related antigens), using h u m a n red cells of c o m m o n and rare Rh phenotypes in an attempt to identify their specificity and/or usefulness as reagents. The report is divided into 3 sections : section I anti-D antibodies, section II other Rh specifitities, section III <>antibodieS.
ANTI-D
21 antibodies, all human. Seven antibodies agglutinated D+ cells (Table I). These antibodies were assum e d to be IgM, the other 1 antibodies were assumed to be IgG. For 6 of the 7 IgM
TABLE 1
Monoclonal anti-D screening and cross-blocking tests. blocked number IgM
Sal
score
Pap
titre
IAT
by of
antibodies
score
titre
6WI
43
160
62
320
45
160
4
8
6W4
52
64
52
64
50
32
3
0 11
6W5
44
32
56
64
43
32
5
6
3
13W3
51
32
99
1024
104
1024
I
7
6
16W6
71
128
83
256
71
128
5
7
2
score
titre
tb pb nb 2
24W4
43
80
55
160
31
40
I
3 10
26W5
55
64
73
128
29
16
4
5
5
168
TIPPEIT P
blocks
for
number
of
+ IgG
IAT
Pap
antibodies
score
titre
6W3
44
160
6W6
42
80
56
6W7
40
80
38
i
6W8
67
320
64
IOW1
30
8
74
10W2
35
80
10W3
31
score I titre 62 320
tb p b nb 0
5 2
160
0
3
4
160
0
3
4
I
320
I
3
3
=
128
0
5
2
61
320
7
0
0
8
37
16
0
I
6
IOW4
49
160
46
160
4
2
I
IOW5
76
256
81
256
I
4
2
16W5
93
512
97
1024
5
2
0
23W3
52
32
55
64
0
I
6
26W6
74
128
100
512
5
2
0
26W7
84
256
74
128
0
4
3
26W8
69
640
49
160
0
I
6
* using
polyspecific
+ using
anti-human
anti-human
globulin
reagent
IgG r e a g e n t
Sal = a g g l u t i n a t i o n
of u n t r e a t e d
Pap = agglutination
of p a p a i n
IAT = a n t i g l o b u l i n
test
using
cells
treated
read microscopically cells
untreated
read microscopically
cells
in
tubes
with
centrifugation. tb = t o t a l l y
blocked,
pb = p a r t i a l l y
blocked,
nb = not
blocked
l
anti-D scores and titres for papain-treated cells were very similar to those of untreated cells. Only 13 W 3 gave notably stronger reactions with papain treated cells than untreated cells. Although the IAT results for the IgM antibodies often did not depend on the addition of antiglobulin reagent, they are included to show whether the antibody eluted during the washing. Considering the 14 IgG anti-D antibodies, very similar results were found for the two techniques used (Table I). IAT was more efficient than agglutination of papain treated cells for 3 antibodies (10 W 1, 10 W 2 and 26 W 6) and less efficient for 26 W 7 ans 26 W 8.
169
MONOCLONAL R H ANTIBODIES TABLE II
Monoclonal anti-D against cells of unusual D phenotype. Category
Saline
IVa
IVb
D
Va
Weak
VI
D
AR
BC
Ko .
Rh:33
6WI
+
+
-
w
.
6W4
+
+
-
+
-
6W5
+
+
13W3
+
+
+/-
-
16W6
+
+
+/-
-
w
.
24W4
+
+
+
-
+
+
-
26W5
+
+
+
-
+
+
.
.
.
+
.
i
ii
Du
D u
_
_
-
_
.
-
.
.
.
+ .
.
.
Papain 6WI
+
+
+
-
6W4
+
+
+
-
6W5
+
+
-
-
13W3
+
+
+
-
16W6
+
+
+
-
24W4
+
+
+
-
26W5
+
+
+
-
6W3
+
+
+/-
w
+
+/-
W
6W6
+
6W7
w/-
w
-
+
+
-
--
-
+/W
+
--
+
W
-
W
+
-
-
+
-
W/-
+
-
+
-
_
+
+
-
-
+/-
+
+
+
-
+
i
ii
+
+
-
+
Du
D u
-
+ / -
-
w
6W8
+
+
+
-
IOWI
+
+
+
-
IOW2
+
+
+
-
IOW3
+
-
+
+
IOW4
+
+
+
-
IOW5
+
+
+
-
16W5
+
+
+
+/w
23W3
-
-
+
-
26W6
+
+
+
-
26W7
+
+
+
-
26W8
+
+
+
-
2
2
4
4
I
+ +/w
+
.
.
.
.
-
-
.
.
.
+
+
+
-
+
+
.
+
-
+
-
+/_
.
+
w
IAT
.
+/w
-
-
_
-
W/-
-
. +w
.
+
+ +
-
-
w/-
+
+
+
-
-
+/w
-
+
+
+
-
-
-
w/-
-
+
-
+
+
-
w
-
+
-
+I-
~/w
-
_
+
+
-
-
+
+
+
+
+
+
+
+
+
+
-
W/-
+
-
W
L ,_
2
3
No.of samples
We
are
unusual
grateful D
phenotypes.
to
many
4
colleagues
who
have
sent
samples
with
17 0
TIPPETFP. The results of blocking tests of IgM anti-D by IgG anti-D were disappointing
(Table I). Only 6 W 4 gave a clear pattern ; it was totally inhibited by 3 antibodies (10 W 2, 16 W 5, 26 W 6) and not inhibited at all by the other 11 IgG antibodies. No definite patterns were observed for the other IgM antibodies. Some were difficult to inhibit (13W3, 2 4 W 4 ) and others relatively easy to inhibit (6W 1, 6 W 5 , 16 W 6). The IgG antibody 10 W 2 inhibited all IgM reagents, 16 W 5 and 26 W 6 were good ~ blockers ~ but 10 W 3, 23 W 3 and 26 W 8 were very inefficient ~ blockers ~. The anti-D antibodies could be used to distinguish category Dw, Dv and Du cells (Table II). All antibodies reacted with Dm (2 Dm and 1 Dmcsamples) and Dw (3 samples) cells. Only 2 antibodies 10 W 3 and 16 W 5 detected Du samples. Most antibodies, 16 of the 21 tested, reacted with all categories except Dw. Three other anfi-D were particularly useful in distinguishing the different categories : 6 W 5 did not react with Dv or Dw cells ; 6 W 7 did not react with Dv or Dw cells and only reacted weakly with Dw ; 23 W 3 did not react with Dr¢ or Dw but reacted with Dv cells. 10 W 3 reacted with Drva cells but not with D~ cells. The results for papain treated cells are presented because, for D category cells, these were stronger than IAT results, especially for Dv cells. The strength of reactions of some antibodies varied from positive or weakly positive to negative for cells from members of the same category or subdivision of a category. The order of strenght of reactions was the same as that found with polyclonal anti-D sera, Some of the IgM anti-D agglutinated papain treated cells (Dv, Rh : 33, AR) although they did not agglutinate untreated cells (Table II). Antibodies which appeared to be similar in their reactions with D category cells gave different reactions with other unusual D phenotypes, weak D antigens (AR, BC, Ko), Du samples and c (D)(e)/cde Rh : 33 samples. The Du samples fall into two types those (the stronger ones as judged by polyclonal reagents) that are more easily detected by IAT than when papain treated and those which are only detected when papain treated (Table II). The variation in the pattern of reactions with the 21 anti-D antibodies suggests that there are many types of Du, as is well known from the observations with polyclonal anti-D. The ability to detect Du and weak D samples does not appear to reflect quantitative differences in the anti-D antibodies. It is interesting that 24 W 4 agglutinates R h : 3 3 cells, and that more of the 21 anti-D are capable of detecting the weak D associated with Rh 33 than detect Dv~ antigens. None of the anti-D reacted with Rh : 33 cells by IAT. None of the IgG anti-D, nor cocktails of them, agglutinated saline suspensions of homozygous - D or .D. cells.
OTHER Rh SPECIFICITIES Anti-c. 1 antibody 26 W 9, human It did not agglutinate untreated c+ cells : it reacted by IAT and agglutinated papain-treated cells. Titration scores and titres for 26 W 9 with c+ cells were: CDe/cde (papain score 48 titre 160 ; IAT 41, 160), CDe/cDE (pap 47, 160 ; IAT 44, 80), CDE/cDE (pap 48, 160 ; IAT 40, 80), cde/cde (pap 44, 160 ; IAT 58, 320), 26 W 9 did not react with CDe/CDe, C'~De/CDe, CDe/CDE, CDE/CDE, - D - , .D., Rhmod,Rhnullcells, or with thawed c+Rh : 26 cells.
MONOCLONAL RH ANTIBODIES
171 TABLE III
Comparison of stTengths of anti-E, - e and -G. t
Anti-E 16W7 pap score
titre
16W8 sal
16W8 pap
score titre
26WI0
score titre
score
IAT
26W15
tizre
score
pap titre
R2R 2
68
32
77
64
86
128
R1R 2 r"r
69
32
80
64
86
68
32
77
64
87
ryr
70
32
73
64
85
128
99
256
21
2
Rzr
50
16
49
32
70
64
90
256
21
2
29W]
IAT
98
256
23
4
128
98
512
21
2
128
101
512
23
4
Anti-e 19W6
IAT
score titre
25W1
IAT
25W2 IAT
score titre~score
RIR I
58
160
45
R ]r
51
i60
42
104 104 104
rr
43
80
42
RTR2
50
160
4]
r"r
35
45
104
104
45
104
104
46
i04
39
104
39
104
42
10
2
0
7
10
10
8
10
8
10
4
0
R2R z
18
20
10
19W6 pap
25WI
94
104
43
80
R2R 2
score
titre 105
105
44
score 51
47
104
7
R]R I
titre
1o 4
titre 5120
pap
25W2
pap
score titre Iscore 54 + >105
29WI
~l~_e score
52 +
pap titre
>I0-"
52 +
>105
>105
50 +
>] 05
rr
95
2560
49
105
54
R2R 2
80
2560
24
102
44
104
34
!03
R2R z
86
2560
37
104
45
104
38
104
Anti-G 23W2 pap score
titre'
23W2 score
IAT
26W11
titre score
pap
26W11
IAT
titre
score
titre
Ro
25
2
18
2
94
256
78
64
r'r
28
2
16
2
98
256
85
128
Tests
against
and positive Negative
samples with
with both
with weak G antigens.
26W11:
R2VI
r (I],,
23W2 and 26W11:
Negative
R2U r (3),
R2Ur"(1).
All antibodies negative with Rhnull cells.
with
23W2
ri'r G+
(2).
172
TIPPETF R
TABLE IV
Summary of results for anti-'Rh' antibodies. 5W2 Titre
against
Saline
rr 103
IAT papain
13W2
19W3
19W4
20W7
27W4
35W2
100
20
105
cells I
0
200
103
I
104
800
10
>106
>105
104
80
104
400
105
>106
>105
-
-
+
Cells Rhnull
U+
w
-
+
w
+
-
w
+
+
+
w
-
w
W
Rhnull
U-
w
-
-
Rhmo d U+
w
-
-
Rhmo d U-
w
-
-
-
-D-
w
+
+
+
+
+
+
cwD -
w
+w
+
+
+
+
+
.D.
w
-
+
+
+
+
+
LW(a-bLW(a-b+
All
+
+
-
+
+
÷w
antibodies
cord,
U-,
Effect
of
positive
with
M k M k,
h r S_ h r B - .
enzyme
treatment
+w
the
+
+
+
+
+
+
following
cells:
D+
cord,
D-
of c e l l s
papain
se
e
e
e/se
e/u
u
trypsin
se
e
u
se/u u
e/u
e/u
u
pronase
se
e
d
d
e/se
d
u
I
e = elevated, d = very
se = s l i g h t l y
depressed/abolished.
elevated,
u : unchanged,
Saline/IAT
where
different
effects
observed We
are
grateful
phenotype.
to many
colleagues
for
sending
cells
of
rare
Rh
173
MONOCLONAL R H ANTIBODIES
Anti-E. 4 antibodies, human.
(Table III)
Only one antibody, 16 W 8, agglutinated E+ cells. Three antibodies, 16 W 7, 16 W 8, 25 W 15 did not react with E negative cells used (C"I)e]CDe, CDe/CDe, CDe/cde, Cde/cde, cde/cde) by agglutination, papain or IAT techniques. 26 W 10 did not react with these samples by agglutination or IAT methods but did react with papain treated cde]cde, CDe/cde and Cde/cde cells. Two antibodies 26 W 10 (IAT) and 26 W 15 (papain) reacted with EW+cells ; 16 W 7 and 16 W 8 did not react with EW+ cells. A n t i - e . 4 a n t i b o d i e s , m u r i n e . (Table III) None of these antibodies agglutinated e+ cells, Three antibodies 25 W 1, 25 W 2 and 29 W 1 behaved as anti-e, if used suitably diluted, by IAT. Although 19 W 6 failed to react with cDE/cDE cells by IAT, it gave strong positive reactions with CDE/CDE and cDE]CDE cells. None of these antibodies behaved as anti-e when papain-treated cells were used (Table III). Anti-G. 2 a n t i b o d i e s , h u m a n .
(Table III)
26 W 11 detected more weak G antigens than 23 W 2 ; this may be due to a quantitative rather than a qualitative difference.
ANTI-~ R H ~ 7 antibodies, murine. (Table IV) The rare phenotypes used are summarized in Table IV. All antibodies were tested against D+ cells blocked with h u m a n anti-LW~, -LW~b, or -D. The antibodies had been called anti-~ Rh ~ because they did not react, or only reacted weakly, with RH~ 1 cells. Some antibodies were very strong, and an end point was not reached in the titrations with rr cells (Table IV). Cells of rare phenotype showed 19 W 3 and 19 W 4 to be anti- LW~b.The effect of enzymes and blocking tests confirmed that 19 W 3 and 19 W 4 were anti-LW~b. Rhnun and Rhmod U-I-cells reacted with 27 W 4 and Rhn~ and Rh~oa U- cells did not react with 27 W 4 (Table lid.
167
Monoclonal Rh antibodies by P. Tippett and C. Lomas MRC Blood Group Unit, Wolfson House, 4 Stephenson Way, London NW1 2HE, U.K.
The Rh antibodies were studied by m a n u a l serological techniques (see Daniels' report on Kell related antigens), using h u m a n red cells of c o m m o n and rare Rh phenotypes in an attempt to identify their specificity and/or usefulness as reagents. The report is divided into 3 sections : section I anti-D antibodies, section II other Rh specifitities, section III <
ANTI-D
21 antibodies, all human. Seven antibodies agglutinated D+ cells (Table I). These antibodies were assum e d to be IgM, the other 1 antibodies were assumed to be IgG. For 6 of the 7 IgM
TABLE 1
Monoclonal anti-D screening and cross-blocking tests. blocked number IgM
Sal
score
Pap
titre
IAT
by of
antibodies
score
titre
6WI
43
160
62
320
45
160
4
8
6W4
52
64
52
64
50
32
3
0 11
6W5
44
32
56
64
43
32
5
6
3
13W3
51
32
99
1024
104
1024
I
7
6
16W6
71
128
83
256
71
128
5
7
2
score
titre
tb pb nb 2
24W4
43
80
55
160
31
40
I
3 10
26W5
55
64
73
128
29
16
4
5
5
168
TIPPEIT P
blocks
for
number
of
+ IgG
IAT
Pap
antibodies
score
titre
6W3
44
160
6W6
42
80
56
6W7
40
80
38
i
6W8
67
320
64
IOW1
30
8
74
10W2
35
80
10W3
31
score I titre 62 320
tb p b nb 0
5 2
160
0
3
4
160
0
3
4
I
320
I
3
3
=
128
0
5
2
61
320
7
0
0
8
37
16
0
I
6
IOW4
49
160
46
160
4
2
I
IOW5
76
256
81
256
I
4
2
16W5
93
512
97
1024
5
2
0
23W3
52
32
55
64
0
I
6
26W6
74
128
100
512
5
2
0
26W7
84
256
74
128
0
4
3
26W8
69
640
49
160
0
I
6
* using
polyspecific
+ using
anti-human
anti-human
globulin
reagent
IgG r e a g e n t
Sal = a g g l u t i n a t i o n
of u n t r e a t e d
Pap = agglutination
of p a p a i n
IAT = a n t i g l o b u l i n
test
using
cells
treated
read microscopically cells
untreated
read microscopically
cells
in
tubes
with
centrifugation. tb = t o t a l l y
blocked,
pb = p a r t i a l l y
blocked,
nb = not
blocked
l
anti-D scores and titres for papain-treated cells were very similar to those of untreated cells. Only 13 W 3 gave notably stronger reactions with papain treated cells than untreated cells. Although the IAT results for the IgM antibodies often did not depend on the addition of antiglobulin reagent, they are included to show whether the antibody eluted during the washing. Considering the 14 IgG anti-D antibodies, very similar results were found for the two techniques used (Table I). IAT was more efficient than agglutination of papain treated cells for 3 antibodies (10 W 1, 10 W 2 and 26 W 6) and less efficient for 26 W 7 ans 26 W 8.
169
MONOCLONAL R H ANTIBODIES TABLE II
Monoclonal anti-D against cells of unusual D phenotype. Category
Saline
IVa
IVb
D
Va
Weak
VI
D
AR
BC
Ko .
Rh:33
6WI
+
+
-
w
.
6W4
+
+
-
+
-
6W5
+
+
13W3
+
+
+/-
-
16W6
+
+
+/-
-
w
.
24W4
+
+
+
-
+
+
-
26W5
+
+
+
-
+
+
.
.
.
+
.
i
ii
Du
D u
_
_
-
_
.
-
.
.
.
+ .
.
.
Papain 6WI
+
+
+
-
6W4
+
+
+
-
6W5
+
+
-
-
13W3
+
+
+
-
16W6
+
+
+
-
24W4
+
+
+
-
26W5
+
+
+
-
6W3
+
+
+/-
w
+
+/-
W
6W6
+
6W7
w/-
w
-
+
+
-
--
-
+/W
+
--
+
W
-
W
+
-
-
+
-
W/-
+
-
+
-
_
+
+
-
-
+/-
+
+
+
-
+
i
ii
+
+
-
+
Du
D u
-
+ / -
-
w
6W8
+
+
+
-
IOWI
+
+
+
-
IOW2
+
+
+
-
IOW3
+
-
+
+
IOW4
+
+
+
-
IOW5
+
+
+
-
16W5
+
+
+
+/w
23W3
-
-
+
-
26W6
+
+
+
-
26W7
+
+
+
-
26W8
+
+
+
-
2
2
4
4
I
+ +/w
+
.
.
.
.
-
-
.
.
.
+
+
+
-
+
+
.
+
-
+
-
+/_
.
+
w
IAT
.
+/w
-
-
_
-
W/-
-
. +w
.
+
+ +
-
-
w/-
+
+
+
-
-
+/w
-
+
+
+
-
-
-
w/-
-
+
-
+
+
-
w
-
+
-
+I-
~/w
-
_
+
+
-
-
+
+
+
+
+
+
+
+
+
+
-
W/-
+
-
W
L ,_
2
3
No.of samples
We
are
unusual
grateful D
phenotypes.
to
many
4
colleagues
who
have
sent
samples
with
17 0
TIPPETFP. The results of blocking tests of IgM anti-D by IgG anti-D were disappointing
(Table I). Only 6 W 4 gave a clear pattern ; it was totally inhibited by 3 antibodies (10 W 2, 16 W 5, 26 W 6) and not inhibited at all by the other 11 IgG antibodies. No definite patterns were observed for the other IgM antibodies. Some were difficult to inhibit (13W3, 2 4 W 4 ) and others relatively easy to inhibit (6W 1, 6 W 5 , 16 W 6). The IgG antibody 10 W 2 inhibited all IgM reagents, 16 W 5 and 26 W 6 were good ~ blockers ~ but 10 W 3, 23 W 3 and 26 W 8 were very inefficient ~ blockers ~. The anti-D antibodies could be used to distinguish category Dw, Dv and Du cells (Table II). All antibodies reacted with Dm (2 Dm and 1 Dmcsamples) and Dw (3 samples) cells. Only 2 antibodies 10 W 3 and 16 W 5 detected Du samples. Most antibodies, 16 of the 21 tested, reacted with all categories except Dw. Three other anfi-D were particularly useful in distinguishing the different categories : 6 W 5 did not react with Dv or Dw cells ; 6 W 7 did not react with Dv or Dw cells and only reacted weakly with Dw ; 23 W 3 did not react with Dr¢ or Dw but reacted with Dv cells. 10 W 3 reacted with Drva cells but not with D~ cells. The results for papain treated cells are presented because, for D category cells, these were stronger than IAT results, especially for Dv cells. The strength of reactions of some antibodies varied from positive or weakly positive to negative for cells from members of the same category or subdivision of a category. The order of strenght of reactions was the same as that found with polyclonal anti-D sera, Some of the IgM anti-D agglutinated papain treated cells (Dv, Rh : 33, AR) although they did not agglutinate untreated cells (Table II). Antibodies which appeared to be similar in their reactions with D category cells gave different reactions with other unusual D phenotypes, weak D antigens (AR, BC, Ko), Du samples and c (D)(e)/cde Rh : 33 samples. The Du samples fall into two types those (the stronger ones as judged by polyclonal reagents) that are more easily detected by IAT than when papain treated and those which are only detected when papain treated (Table II). The variation in the pattern of reactions with the 21 anti-D antibodies suggests that there are many types of Du, as is well known from the observations with polyclonal anti-D. The ability to detect Du and weak D samples does not appear to reflect quantitative differences in the anti-D antibodies. It is interesting that 24 W 4 agglutinates R h : 3 3 cells, and that more of the 21 anti-D are capable of detecting the weak D associated with Rh 33 than detect Dv~ antigens. None of the anti-D reacted with Rh : 33 cells by IAT. None of the IgG anti-D, nor cocktails of them, agglutinated saline suspensions of homozygous - D or .D. cells.
OTHER Rh SPECIFICITIES Anti-c. 1 antibody 26 W 9, human It did not agglutinate untreated c+ cells : it reacted by IAT and agglutinated papain-treated cells. Titration scores and titres for 26 W 9 with c+ cells were: CDe/cde (papain score 48 titre 160 ; IAT 41, 160), CDe/cDE (pap 47, 160 ; IAT 44, 80), CDE/cDE (pap 48, 160 ; IAT 40, 80), cde/cde (pap 44, 160 ; IAT 58, 320), 26 W 9 did not react with CDe/CDe, C'~De/CDe, CDe/CDE, CDE/CDE, - D - , .D., Rhmod,Rhnullcells, or with thawed c+Rh : 26 cells.
MONOCLONAL RH ANTIBODIES
171 TABLE III
Comparison of stTengths of anti-E, - e and -G. t
Anti-E 16W7 pap score
titre
16W8 sal
16W8 pap
score titre
26WI0
score titre
score
IAT
26W15
tizre
score
pap titre
R2R 2
68
32
77
64
86
128
R1R 2 r"r
69
32
80
64
86
68
32
77
64
87
ryr
70
32
73
64
85
128
99
256
21
2
Rzr
50
16
49
32
70
64
90
256
21
2
29W]
IAT
98
256
23
4
128
98
512
21
2
128
101
512
23
4
Anti-e 19W6
IAT
score titre
25W1
IAT
25W2 IAT
score titre~score
RIR I
58
160
45
R ]r
51
i60
42
104 104 104
rr
43
80
42
RTR2
50
160
4]
r"r
35
45
104
104
45
104
104
46
i04
39
104
39
104
42
10
2
0
7
10
10
8
10
8
10
4
0
R2R z
18
20
10
19W6 pap
25WI
94
104
43
80
R2R 2
score
titre 105
105
44
score 51
47
104
7
R]R I
titre
1o 4
titre 5120
pap
25W2
pap
score titre Iscore 54 + >105
29WI
~l~_e score
52 +
pap titre
>I0-"
52 +
>105
>105
50 +
>] 05
rr
95
2560
49
105
54
R2R 2
80
2560
24
102
44
104
34
!03
R2R z
86
2560
37
104
45
104
38
104
Anti-G 23W2 pap score
titre'
23W2 score
IAT
26W11
titre score
pap
26W11
IAT
titre
score
titre
Ro
25
2
18
2
94
256
78
64
r'r
28
2
16
2
98
256
85
128
Tests
against
and positive Negative
samples with
with both
with weak G antigens.
26W11:
R2VI
r (I],,
23W2 and 26W11:
Negative
R2U r (3),
R2Ur"(1).
All antibodies negative with Rhnull cells.
with
23W2
ri'r G+
(2).
172
TIPPETF R
TABLE IV
Summary of results for anti-'Rh' antibodies. 5W2 Titre
against
Saline
rr 103
IAT papain
13W2
19W3
19W4
20W7
27W4
35W2
100
20
105
cells I
0
200
103
I
104
800
10
>106
>105
104
80
104
400
105
>106
>105
-
-
+
Cells Rhnull
U+
w
-
+
w
+
-
w
+
+
+
w
-
w
W
Rhnull
U-
w
-
-
Rhmo d U+
w
-
-
Rhmo d U-
w
-
-
-
-D-
w
+
+
+
+
+
+
cwD -
w
+w
+
+
+
+
+
.D.
w
-
+
+
+
+
+
LW(a-bLW(a-b+
All
+
+
-
+
+
÷w
antibodies
cord,
U-,
Effect
of
positive
with
M k M k,
h r S_ h r B - .
enzyme
treatment
+w
the
+
+
+
+
+
+
following
cells:
D+
cord,
D-
of c e l l s
papain
se
e
e
e/se
e/u
u
trypsin
se
e
u
se/u u
e/u
e/u
u
pronase
se
e
d
d
e/se
d
u
I
e = elevated, d = very
se = s l i g h t l y
depressed/abolished.
elevated,
u : unchanged,
Saline/IAT
where
different
effects
observed We
are
grateful
phenotype.
to many
colleagues
for
sending
cells
of
rare
Rh
173
MONOCLONAL R H ANTIBODIES
Anti-E. 4 antibodies, human.
(Table III)
Only one antibody, 16 W 8, agglutinated E+ cells. Three antibodies, 16 W 7, 16 W 8, 25 W 15 did not react with E negative cells used (C"I)e]CDe, CDe/CDe, CDe/cde, Cde/cde, cde/cde) by agglutination, papain or IAT techniques. 26 W 10 did not react with these samples by agglutination or IAT methods but did react with papain treated cde]cde, CDe/cde and Cde/cde cells. Two antibodies 26 W 10 (IAT) and 26 W 15 (papain) reacted with EW+cells ; 16 W 7 and 16 W 8 did not react with EW+ cells. A n t i - e . 4 a n t i b o d i e s , m u r i n e . (Table III) None of these antibodies agglutinated e+ cells, Three antibodies 25 W 1, 25 W 2 and 29 W 1 behaved as anti-e, if used suitably diluted, by IAT. Although 19 W 6 failed to react with cDE/cDE cells by IAT, it gave strong positive reactions with CDE/CDE and cDE]CDE cells. None of these antibodies behaved as anti-e when papain-treated cells were used (Table III). Anti-G. 2 a n t i b o d i e s , h u m a n .
(Table III)
26 W 11 detected more weak G antigens than 23 W 2 ; this may be due to a quantitative rather than a qualitative difference.
ANTI-~ R H ~ 7 antibodies, murine. (Table IV) The rare phenotypes used are summarized in Table IV. All antibodies were tested against D+ cells blocked with h u m a n anti-LW~, -LW~b, or -D. The antibodies had been called anti-~ Rh ~ because they did not react, or only reacted weakly, with RH~ 1 cells. Some antibodies were very strong, and an end point was not reached in the titrations with rr cells (Table IV). Cells of rare phenotype showed 19 W 3 and 19 W 4 to be anti- LW~b.The effect of enzymes and blocking tests confirmed that 19 W 3 and 19 W 4 were anti-LW~b. Rhnun and Rhmod U-I-cells reacted with 27 W 4 and Rhn~ and Rh~oa U- cells did not react with 27 W 4 (Table lid.