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Antibody formation during the development of acute graft-versus-host disease after small-bowel transplantation
ELSEVIER
Anti body Graft-Versus-Host
During the Development of Acute Disease After Small-Bowel Transplantation
C. Johnsson, M. Bengtsson, and G. Tufveson
A
LTHOUGH a rare event, graft-versus-host disease (GVHD) has been diagnosed in small-bowel graft recipients, and has, in some cases, contributed to patient death. T lymphocytes of donor origin are crucial for the induction of GVHD.‘y* However, little is known about the effector mechanisms that lead to destruction of recipient tissue. The present study was undertaken to study whether humoral immunity is involved in acute GVHD. MATERIALS AND METHODS
Heterotopic small-bowel transplantations were performed using Lewis rats as donors and (Lewis xBN)Fl-hybrids as recipients. On day 3, 5, or 7 the animals were killed (3 to 4 each time). The presence of lymphocytotoxic antibodies in sera was analysed using a standard microcytotoxic NIH technique with spleen cells from BN, Lewis, and WistarKyoto rats as target. The existence of IgG and IgM antibodies in the liver was examined by immunofluorescent staining. RESULTS AND DISCUSSION
Mild clinical symptoms of GVHD, such as redness of ears and paws, were observed on day 7 (Table 1). No severe signs of GVHD were seen during the observation period; untreated GVHD in this strain combination is invariably lethal, with a median recipient survival time of 14 days.3 Antibodies directed against BN spleen cells started to appear in recipient serum on day 5 posttransplantation (Table 1) and a slightly further increase was seen on day 7. No antibodies directed against either donor (Lewis) or third-party (Wistar/Kyoto) spleen cells were detectable. Concomitantly with the rise in antibody-titer in serum, IgM antibodies became visible in the liver (Table 1). The liver is one of the target organs for GVHD in which early signs of the disease can be detected.4 No IgM antibodies were seee in livers from untreated, nontransplanted animals. In contrast, IgG antibodies were found in liver biopsies from untreated, nontransplanted rats, and no clear quantitative difference could be seen during the development of GVBD.
Q 1997 by Elsevier Science inc. 655 Avenue of the Americas, New York, NY 10010
Table 1. Clinical Appearance and Presence of Antibodies in Serum and Liver After Semisyngeneic Snail-Bowel Transplantation
Clinical signs of GVHD Recipient-reactive antibodies
Day 0
Day 3
Day 5
-
_
-
-
-
-
-Ii
++/i-++
Day 7
+ +++
in serum
IgM antibodies in the liver
++
++
Clinicalsigns of GVHD were graded on a scale from - (no signs of GVHD) to + + + (severe signs of GVHD).The numbers of cytotoxic antibodies in serum were graded from - (0% to 15% cell death) to + +++ (75”%to 100% cell death). The numbers of IgM antibodies in the liver were graded arbitrarily from - (no deposition of IgM) to + + + (pronounced deposition of IgM).
In conclusion, we have demonstrated high titers of recipient-reactive antibodies in rats during the development of acute GVHD. It can be assumed that these antibodies contribute to the disease, although this remains to be clearly demonstrated. Furthermore, antibodies in sera appeared before any clinical symptoms of GVHD could be seen. Thus, we suggest that determination of recipient-reactive antibodies in serum could function as a diagnostic tool in small-bowel graft recipients. REFERENCES 1. Kirkman RL, Lear PA, Madara JL, et al: Surgery 96:280,1984 2. Wallander J, L.&kgren G, Tufveson G: Transplant Proc 21:2896, 1989 3. Johnsson C, Tufveson G: Transplantation 57:289, 1994 4. Johnsson C, Gannedahl G, Scheynius A, et al: Transplantation 59:1524, 1995 From Departments of Transplantation Surgery (C.J., G.T.) and Clinical Immunology and Transfusion Medicine (M.B.), University Hospital, Uppsala, Sweden. This work was supported by the Swedish Medical Research Council. Address reprint requests to Cecilia Johnsson, Dept. of Transplantation Surgery, University Hospital, S-751 85 Uppsala, Sweden.
0041-l 345/97&l 7.00 PII SO041 -1345(96)00423-X
699
Transplantation
Proceedings, 29, 699 (1997)
Anti body Graft-Versus-Host
During the Development of Acute Disease After Small-Bowel Transplantation
C. Johnsson, M. Bengtsson, and G. Tufveson
A
LTHOUGH a rare event, graft-versus-host disease (GVHD) has been diagnosed in small-bowel graft recipients, and has, in some cases, contributed to patient death. T lymphocytes of donor origin are crucial for the induction of GVHD.‘y* However, little is known about the effector mechanisms that lead to destruction of recipient tissue. The present study was undertaken to study whether humoral immunity is involved in acute GVHD. MATERIALS AND METHODS
Heterotopic small-bowel transplantations were performed using Lewis rats as donors and (Lewis xBN)Fl-hybrids as recipients. On day 3, 5, or 7 the animals were killed (3 to 4 each time). The presence of lymphocytotoxic antibodies in sera was analysed using a standard microcytotoxic NIH technique with spleen cells from BN, Lewis, and WistarKyoto rats as target. The existence of IgG and IgM antibodies in the liver was examined by immunofluorescent staining. RESULTS AND DISCUSSION
Mild clinical symptoms of GVHD, such as redness of ears and paws, were observed on day 7 (Table 1). No severe signs of GVHD were seen during the observation period; untreated GVHD in this strain combination is invariably lethal, with a median recipient survival time of 14 days.3 Antibodies directed against BN spleen cells started to appear in recipient serum on day 5 posttransplantation (Table 1) and a slightly further increase was seen on day 7. No antibodies directed against either donor (Lewis) or third-party (Wistar/Kyoto) spleen cells were detectable. Concomitantly with the rise in antibody-titer in serum, IgM antibodies became visible in the liver (Table 1). The liver is one of the target organs for GVHD in which early signs of the disease can be detected.4 No IgM antibodies were seee in livers from untreated, nontransplanted animals. In contrast, IgG antibodies were found in liver biopsies from untreated, nontransplanted rats, and no clear quantitative difference could be seen during the development of GVBD.
Q 1997 by Elsevier Science inc. 655 Avenue of the Americas, New York, NY 10010
Table 1. Clinical Appearance and Presence of Antibodies in Serum and Liver After Semisyngeneic Snail-Bowel Transplantation
Clinical signs of GVHD Recipient-reactive antibodies
Day 0
Day 3
Day 5
-
_
-
-
-
-
-Ii
++/i-++
Day 7
+ +++
in serum
IgM antibodies in the liver
++
++
Clinicalsigns of GVHD were graded on a scale from - (no signs of GVHD) to + + + (severe signs of GVHD).The numbers of cytotoxic antibodies in serum were graded from - (0% to 15% cell death) to + +++ (75”%to 100% cell death). The numbers of IgM antibodies in the liver were graded arbitrarily from - (no deposition of IgM) to + + + (pronounced deposition of IgM).
In conclusion, we have demonstrated high titers of recipient-reactive antibodies in rats during the development of acute GVHD. It can be assumed that these antibodies contribute to the disease, although this remains to be clearly demonstrated. Furthermore, antibodies in sera appeared before any clinical symptoms of GVHD could be seen. Thus, we suggest that determination of recipient-reactive antibodies in serum could function as a diagnostic tool in small-bowel graft recipients. REFERENCES 1. Kirkman RL, Lear PA, Madara JL, et al: Surgery 96:280,1984 2. Wallander J, L.&kgren G, Tufveson G: Transplant Proc 21:2896, 1989 3. Johnsson C, Tufveson G: Transplantation 57:289, 1994 4. Johnsson C, Gannedahl G, Scheynius A, et al: Transplantation 59:1524, 1995 From Departments of Transplantation Surgery (C.J., G.T.) and Clinical Immunology and Transfusion Medicine (M.B.), University Hospital, Uppsala, Sweden. This work was supported by the Swedish Medical Research Council. Address reprint requests to Cecilia Johnsson, Dept. of Transplantation Surgery, University Hospital, S-751 85 Uppsala, Sweden.
0041-l 345/97&l 7.00 PII SO041 -1345(96)00423-X
699
Transplantation
Proceedings, 29, 699 (1997)