Exeley Inc.

Reis, Marciana D.

Letters to the Editors: Response to Dr. Apfalroth: Ferndandes JR, et al.

Petz, Lawrence D.

BOOK REVIEW: The Autoimmune Diseases

Fields, Angela C.

Grindon, Alfred J.

Hemochromatosis, iron, and blood donation: a short review

Mallory, Delores

McGinniss, Mary

IN MEMORIUM: Aaron M. Josephson, MD (1924-1999)

Daniels, Geoff

DeLong, E. Nicole

Hare, Virginia

Johnson, Susan T.

LePennec, Pierre-Yves

Marshall, M. Jane

Oliver, Cindy

Spruell, Peggy

I M M U N O H E M A T O L O G Y ,  V O L U M E  1 4 ,  N U M B E R  2 ,  1 9 9 8 49
A new high-frequency red cell antigen has been identified and named
GIL. GIL differs from all high-frequency antigens included in the
International Society of Blood Transfusion classification. There is very
little family information and GIL has not been shown to be an inher-
ited character. Five women with anti-GIL have been found. All had
been pregnant at least twice. Red blood cells of two of the babies gave
positive direct antiglobulin tests, but there were no clinical signs of
hemolytic disease. Anti-GIL may have been responsible for a hemolyt-
ic transfusion reaction and results of monocyte monolayer assays of
two of the anti-GIL suggested a potential to cause destruction of trans-
fused GIL+ RBCs. Immunohematology 1998;14:49–52.
Key Words: blood groups, high-frequency antigens, red
cells, GIL
Antibodies to high-frequency red blood cell (RBC)
antigens can be very important in transfusion medicine.
Some are capable of causing severe immediate or
delayed hemolytic transfusion reactions or hemolytic
disease of the fetus and newborn (HDN). Such antibod-
ies create a particular problem, as compatible blood is
often very difficult to find. Many antibodies to high-
frequency RBC antigens have not been shown to be clin-
ically significant but still cause considerable problems in
blood transfusion immunohematology reference labora-
tories, as they require identification. This procedure is
time consuming, requires extensive testing, and may
delay the provision of blood for transfusion.
The International Society for Blood Transfusion (ISBT)
recognizes 12 antigens of very high frequency that have
not been shown to belong to any blood group system or
collection:Vel, Lan,Ata, Jra,Oka, JMH,Emm,AnWj,Duclos,
PEL,ABTI, and MAM.1–3 Many other high-frequency anti-
gens, which do not fill the criteria necessary for recog-
nition by the ISBT, are known by immunohematology
reference laboratories. One such antigen is GIL.Anti-GIL
was first identified in 19804,5 and several other GIL–
individuals have been found since that time.
Materials and Methods
Serologic methods
Standard serologic tests were used involving both nor-
mal-ionic-strength saline and low-ionic-strength saline,
although the methods used differed in the various labo-
ratories involved in the investigation. Antiglobulin tests
were read in the presence of anti-human IgG or polyspe-
cific anti-human globulin after three or four washes of
the sensitized RBCs. For a two-stage complement test,
RBCs were sensitized in the appropriate antiserum,
mixed with 4mg/mL EDTA, washed, incubated for 20
minutes in fresh inert human AB serum, and washed
again, and then the test was read after addition of anti-
human C4+C3. Eluates were prepared from sensitized
RBCs with an acid-elution kit (Elu-kit II, Gamma
Biologicals, Houston, TX). RBCs were treated with 4%
ficin and with papain, trypsin, α-chymotrypsin, and
dithiothreitol (DTT),as described previously.6–8 IgG sub-
classing was performed by a capillary method.6
Monocyte monolayer assay
Monocyte monolayer assays to predict the clinical sig-
nificance of antibodies were performed by previously
described methods.6,9 Briefly, RBCs sensitized with the
appropriate antibody were incubated with peripheral
blood monocytes in tissue culture chamber slides.After
fixation and staining, the slides were examined micro-
scopically and the percentage of reactive monocytes
was determined.
Immunoblotting
Immunoblotting was carried out as described previ-
ously,10 with membranes prepared in either the pres-
ence or absence of the reducing agent 2-mercap-
toethanol and with an eluate prepared from GIL+ RBCs
sensitized with the Gil antibody.
Results
Propositi
Five propositi have been ascertained through the
presence of an antibody to an unidentified high-fre-
quency RBC antigen,subsequently shown to be anti-GIL.
GIL: a red cell antigen of very high
frequency
G.L. DANIELS, E.N. DELONG,V. HARE, S.T. JOHNSON, P.Y. LEPENNEC, D. MALLORY, M.J. MARSHALL, C. OLIVER,AND P. SPRUELL
G.L. DANIELS ET AL.
Case 1. Gil is a group A white American woman with
no history of blood transfusion. The RBCs of her first
child had a weakly positive direct antiglobulin test
(DAT),but no antibody could be eluted and there was no
clinical sign of HDN. The RBCs of her second baby,
despite reacting with her antibody, had a negative DAT.
The parents of Gil are not consanguineous and her three
siblings are GIL+.
Case 2. Boi is a group O white French woman. Her
antibody was found following a hemolytic transfusion
reaction, which occurred during surgery in a small hos-
pital. Her RBCs had a positive DAT with anti-IgG, but no
eluate was prepared.Soon after the transfusion reaction,
her anti-GIL had a titer of 1,024, which decreased to a
titer of 2 within 3 months. RBCs of her two siblings and
six children reacted with her antibody. No symptoms of
HDN were reported for any of her children.Boi RBCs did
not react with the Gil serum by an antiglobulin test and
two adsorptions of the Gil serum with Boi RBCs did not
remove the antibody, whereas two adsorptions with
GIL+ RBCs removed all antibody. However, a sample of
Boi RBCs taken 13 years later was found to be weakly
reactive with the Gil serum and with an eluate prepared
from GIL+ cells after sensitization with the Hun serum
(see Case 3).The Boi serum (with added soluble A sub-
stance) did not react with Gil RBCs.
Case 3. Hun, a group O white German woman living
in the United States, presented postpartum with an anti-
body to a high-frequency antigen in her serum. RBCs of
the baby had a 3+ DAT, but there was no sign of HDN.
Nine years later, the patient presented again postpartum
(third pregnancy) with an antibody to a high-frequency
antigen, but the baby was unaffected. Hun RBCs did not
react with the Gil serum, and an eluate containing the
antibody of Hun did not react with Gil RBCs.
Case 4. Gou,a group A white woman,presented with
a pelvic infection and possible abdominal mass. She had
a history of four pregnancies and no transfusions. The
Gou serum did not react with Gil RBCs, and Gou RBCs
did not react with the Gil serum, but the Gou serum
reacted weakly with Hun RBCs.
Case 5. Mil, a group O white American woman, pre-
sented with bradycardia when she was 78 years old.
She had received 2 units of packed RBCs 5 weeks ear-
lier when no antibody was detected. She had been
pregnant once. Her RBCs did not react with the Gil
serum and her antibody adsorbed and eluted from
GIL+ RBCs did not react with GIL– RBCs from Gil and
Gou; two adsorptions with GIL– RBCs did not remove
the antibody from the Mil serum. In addition to anti-
GIL, her serum contained anti-P1.
Adsorption tests of the five anti-GIL with selected
GIL+ cells revealed no additional blood group specifici-
ties, with the exception of anti-P1 in the Mil serum.
Serologic characteristics of GIL antigen
GIL has been shown to differ from the following high-
frequency antigens, either by reactivity of Gil RBCs with
the antibodies to those antigens or by the reaction of the
Gil serum with RBCs lacking those antigens: U; Ena;
EnKT;‘N’; EnEP; ENEH; Hr0; Hr; Rh29; Hr
B; Nou; Sec; Dav;
MAR; Lub; Lu3; Lu4; Lu5; Lu6; Lu7; Lu8; Lu11; Lu12; Lu13;
Lu16;Lu17;Lu20;k;Kpb,Ku;Jsb;K11;K12;K13;K14;K16;
K18; K19; Km; K22;Tou; Fy3; Fy5; Jk3; Dib;Wrb;Yta; Sc1;
Sc3;Gya;Hy;Joa;Coa;Co3;LWa;LWab;Ch1;Ch2;Ch3;Ch4;
Ch5;Ch6;Rg1;Rg2;H;Kx;Ge2;Ge3;Ge4;Cra;Tca;Dra;Esa;
IFC;WESb;UMC;Kna;McCa;Sla;Yka;Inb;Csa;I; i;Era;P;LKE;
Vel; Lan;Ata; Jra; Oka; JMH; Emm;AnWj; Duclos; PEL;ABTI;
and MAM.
GIL– RBCs did not react with antibodies to any of the
following low-frequency antigens that have not been
shown to belong to any blood group system: By;Bxa;Tra;
Jra;Toa;Pta;or Jea.Boi RBCs were found to be Rd+,but Gil
and Hun RBCs were subsequently shown to be Rd–.Gou
and Mil RBCs were not tested.
Expression of GIL on RBCs was enhanced by treat-
ment of the cells with papain, ficin, α-chymotrypsin, or
trypsin,and was not affected by DTT treatment. Anti-GIL
reacted slightly less strongly with cord red cells than
with red cells from adults.
Serologic characteristics of three anti-GIL
Anti-GIL did not directly agglutinate GIL+ RBCs but
did react with those RBCs by an indirect antiglobulin
test with polyspecific anti-human globulin or anti-human
IgG.The Boi serum fixed complement as determined by
a two-stage complement technique, whereas the Gil
serum did not.Anti-GIL in the Gil serum was IgG1; that
in the Hun serum was IgG1 plus IgG2.Anti-GIL was not
inhibited by human serum, saliva, Lewis-substance, milk,
urine, or pigeon protein.
Frequency of GIL
The Gil plasma diluted 1:3 was used by a microtiter
method to screen RBCs from 6,300 predominantly white
blood donors and diluted 1:2 by a tube method to screen
RBCs from 2,062 black donors in Milwaukee,WI.The Gil
plasma, diluted 1:2 was used to screen red cells from
2,760 donors in Atlanta, GA, using an Olympus PK-7100
dextran method. RBCs from 101 Asian, 779 black, and
198 Hispanic donors in Jacksonville, FL, were screened
I M M U N O H E M A T O L O G Y ,  V O L U M E  1 4 ,  N U M B E R  2 ,  1 9 9 850
GIL: a high-frequenty antigen
I M M U N O H E M A T O L O G Y ,  V O L U M E  1 4 ,  N U M B E R  2 ,  1 9 9 8 51
with anti-GIL. In Connecticut, RBCs from 14,191 donors
were screened by a microtiter polybrene method with
the Gil serum diluted 1:20. All donors were GIL+, but
RBCs from one of the white Milwaukee donors gave a
much weaker reaction than all others.
Monocyte monolayer assays
Monocyte monolayer assays (MMA) were carried out
with two anti-GIL.With the Gil antibody, the procedure
described in Immunohematology Methods and
Procedures6 was used and results of 22 percent and 23
percent, 32 percent and 27 percent were obtained (nor-
mal range 0–3%). With the Hun serum, the method of
Schanfield et al.9 was used and results of 574 percent
and 765 percent were obtained, where 100 percent rep-
resents the result obtained with a standard IgG1 anti-D.
Both sets of results suggest that these antibodies have a
potential to cause significant RBC destruction of trans-
fused GIL+ RBCs.
Immunoblotting
No bands were detected by immunoblotting with an
anti-GIL eluate derived from the Gil serum, either under
reducing or nonreducing conditions.
Discussion
Antibodies in the sera of five women appeared to be
detecting a new RBC antigen of high frequency.This anti-
gen is different from all of the high-frequency antigens
defined by the ISBT Working Party on Terminology for
Red Cell Surface Antigens1,2 and has been named GIL.
There is a paucity of family information; only the three
siblings of the original GIL– proposita were tested with
anti-GIL and all were GIL+.Consequently, there is no evi-
dence that GIL is an inherited character and so cannot
be included in the ISBT classification. RBCs from 26,391
blood donors from the United States (23,449 white,
2,841 black, and 101 Asian) were tested with anti-GIL
and all were GIL+.
A few anomalous results suggest that all five anti-GIL
may not be of identical specificity. Boi RBCs may have
reacted very weakly with the Gil and Hun sera, although
this was not noticed in the original testing. The Gou
serum reacted very weakly with Hun RBCs.This kind of
heterogeneity among antibodies to high-frequency anti-
gens of similar specificity has been seen with other high-
frequency antigens; for example, PEL.11 Boi RBCs were
positive for the low-frequency antigen Rd (700015),
which is probably associated with the Scianna sys-
tem,12,13 but RBCs of the two other GIL– women (Gil
and Hun) tested with anti-Rd were Rd–.
There is no direct evidence that anti-GIL is clinically
significant. Monocyte monolayer assays were performed
with two examples of anti-GIL and the levels of phago-
cytosis were high enough to suggest that the antibody
had the potential to cause destruction of transfused
incompatible RBCs. One woman, subsequently found to
have anti-GIL, had a hemolytic transfusion reaction, but
no details regarding the severity of this reaction are avail-
able and there is no proof that anti-GIL was responsible.
RBCs from neonates of two of the women with anti-GIL
gave positive DAT reactions, but there was no evidence
of HDN.
The GIL antigen is not detroyed by treatment of the
RBCs with proteolytic enzymes. Immunoblotting with
anti-GIL against membranes derived from GIL+ RBCs
produced no bands. Consequently, very little is known
about the immunochemistry of the GIL antigen.
We will attempt to find further examples of GIL–
individuals and anti-GIL. More family investigations
might demonstrate inheritance of the GIL– phenotype,
permitting GIL to join the 901 Series of high-frequency
antigens in the ISBT classification of RBC cell-surface
antigens.
Acknowledgments
We thank Robert Eckrich, MT(ASCP)SBB, of the
National Reference Laboratory for Blood Group
Serology, Rockville, MD, for monocyte monolayer 
assays and IgG subclassing; William Montgomery,
MT(ASCP)SBB, Florida Georgia Blood Alliance, Jackson-
ville, FL, and Natalie Wilhite, the Blood Center of
Southeastern Wisconsin, Milwaukee, WI, for screening
donors; and Dawn Rumsey, ART, National Reference
Laboratory for Blood Group Serology, Rockville, MD, and
Jill R. Storrey, MS, FIBMS, New York Blood Center, New
York, NY, for serologic testing.
References
1. Daniels GL,Anstee DJ, Cartron JP, et al. Blood group
terminology 1995. From the ISBT Working Party on
Terminology for Red Cell Surface Antigens.Vox Sang
1995;69:265–79.
2. Daniels GL,Anstee DJ, Cartron JP, et al.Terminology
for red cell surface antigens. Makuhari report. Vox
Sang 1996;71:246–8.
3. Montgomery W,Nance S,Kavitsky D,et al.First exam-
ple of hemolytic disease of the newborn (HDN) due
to an antibody to a high incidence antigen present
on multiple cell lines (MCL) (abstract). Transfusion
1997;37(Suppl):41S.
G.L. DANIELS ET AL.
4. Daniels GL. Blood group antigens of high frequency:
a serological and genetical study [Doctoral thesis].
London, England: University of London; 1980.
5. Frederick J, Brendel W, Daniels G, Menitove JE,
Gottschell JL. Gill: investigation of a new high-inci-
dence red cell antigen (abstract).Transfusion 1981;
21(Suppl):613.
6. Beattie K, Crawford M, Mallory D, Mougey R.
Immunohematology methods and procedures.
American National Red Cross, 1993.
7. Daniels G. Effect of enzymes on and chemical modi-
fication of high-frequency red cell antigens.
Immunohematology 1986;8:53–7.
8. Toy EM. Inactivation of high-incidence antigens on
red blood cells by dithiothreitol. Immunohematology
1986;2:57–9.
9. Schanfield MS, Stevens JO, Bauman D.The detection
of clinically significant erythrocyte alloantibodies
using a human mononuclear phagocyte assay.
Transfusion 1981;21:571–6.
10. Daniels GL,Green CA,Okubo Y,et al.SAT,a “new”low
frequency blood group antigen, which may be asso-
ciated with two different MNS variants.Transfus Med
1991;1:39–45.
11. Daniels GL, Simard H, Goldman M, et al. PEL, a “new”
high-frequency red cell surface antigen. Vox Sang
1996;70:31–3.
12. Lewis M, Kaita H, Phillips S, et al.The position of the
Radin blood group locus in relation to other chro-
mosome 1 loci. Ann Hum Genet 1980;44:179–84.
13. Spring FA. Characterization of blood-group-active
erythrocyte membrane glycoproteins with human
antisera.Transfus Med 1993;3:167–78.
Geoff Daniels, PhD, Senior Research Fellow, Bristol
Institute for Transfusion Sciences, Southmead Road,
Bristol, BS10 5ND, UK; E. Nicole DeLong, MS,
MT(ASCP)SBB, Supervisor, Reference and Consultation
Laboratory, American Red Cross Connecticut Region,
Farmington, CT;Virginia Hare, MT(ASCP)SBB,
American Red Cross Blood Services, Southern Region,
Atlanta, GA; Susan T. Johnson, MT(ASCP)SBB,
Supervisor, Immunohematology Reference Laboratory
and Transfusion Service, Milwaukee, WI; Pierre-Yves
LePennec, MD, Centre National de Référence pour les
Groupes Sanguines, Paris Cedex 11, France; Delores
Mallory, American Red Cross, Musser Blood Center,
Technical Services, Philadelphia, PA; M. Jane Marshall,
MT(ASCP)SBB, Florida Georgia Blood Alliance,
Jacksonville, FL; Cindy Oliver, MT(ASCP)SBB, Olney,
MD; and Peggy Spruell, MT(ASCP)SBB, Gamma
Biologicals Inc., Houston, TX.
I M M U N O H E M A T O L O G Y ,  V O L U M E  1 4 ,  N U M B E R  2 ,  1 9 9 852
Attention SBB and BB Students: You are 
eligible for a free one-year subscription to
Immunohematology. Ask your education super-
visor to submit the name and complete address
for each student and the inclusive dates of the
training period to Immunohematology, P.O. Box
40325, Philadelphia, PA 19106.
Attention: State Blood Bank Meeting
Organizers—If you are planning a state meet-
ing and would like copies of Immuno-
hematology for distribution, please contact
Mary H.McGinniss, Managing Editor, 3 months
in advance,by phone or fax at (301) 299-7443.


Not a member yet

Letters to the Editors: Response to Dr. Apfalroth: Ferndandes JR, et al.

BOOK REVIEW: The Autoimmune Diseases

Hemochromatosis, iron, and blood donation: a short review

IN MEMORIUM: Aaron M. Josephson, MD (1924-1999)

GIL: a red cell antigen of very high frequency

A comprehensive IgA service provided by a blood transfusion center

Report on anti-Dib encountered in two Hong Kong Chinese

Letters From the Editors: Immunohematology Is on the World Wide Web

Letter to the Editors: "Those Were the Days"

Letters to the Editors: "Those Were the Days"

4,746

6,096