588 FETAL RESPONSE TO DRUG WITHDRAWAL
SiR,—There have been several reports of withdrawal symp-
well in advance to allow time for drawal of the drug of dependency.
planned
a
gradual with.
in neonates whose mothers were still addicted at delivery. Department of DAVID T. Y. LIU Obstetrics and Gynæcology, These babies appear normal at birth but within 10-36 h ELIZABETH TYLDEN University College Hospital become distressed and require treatment in a neonatal unit.’1 SINA H. TUKEL Medical School, London WC1. It is, therefore, in the infants’ interest that these addicted mothers should be identified and an attempt made to wean them off drugs before delivery. In the past five years 18 known addicts delivered in University College Hospital, London, and ONDINE’S CURSE 11 of these patients were weaned off opiates during pregnancy. readers SIR,Your may interested in a further example It is, however, not generally realised that the fetus may react Ondine’s curse in a young girl. The mythological reference of if or withdrawn adversely drugs are withheld during labour was first applied to three adults who had apnreic periods when rapidly during the antenatal period. Withholding drugs from awake but breathed on command and required mechanical known, or unsuspected, addicts has resulted in a stillbirth and ventilation when asleep.’Ondine, a German water nymph, ina neonatal death in neighbouring hospitals. voked a curse upon her jilted husband so that he would forget to the usually recogDuring labour, fetal distress-not due to breathe (and die) when he fell asleep. nised causes-arising in opiate addicts often appears to be The girl aged 2 years 8 months was admitted with a history relieved by the administration of pethidine or a replacement of recent mild diarrhoea and a family "flu-like" illness. For5 drug such as physeptone. This therapy is now advised in cases she had been intermittently drowsy and prone to sleep. days of opiate dependence. Immediately before admission she became difficult to rouse and We found that during the antenatal period, 24 h urinary cyanosed. During the preceding 9 months the parents had oestrogen production fell for the 48 h after each stepped reducnoticed that "she was blue around the lips" on three occasions tion in drug dosage. Thereafter, the level of maternal oestrogen in the early morning. She was otherwise healthy and had a excretion gradually rose until the drug was reduced again. On normal perinatal history. two occasions drug withdrawal had to be slowed because of Examination revealed a pale, cyanosed child with peripheral precipitate falls in urinary oestriols. limb oedema, puffy face, bounding pulses, and a soft precordial On one occasion, adverse changes in fetal heart-rate were murmur. The cyanosis was reversed by oxygen. Later she observed when external cardiotocography was carried out on became more drowsy with deep cyanosis, papilloedema, and a a patient during an episode of rapid drug withdrawal. A pacranial bruit. tient dependent on pentazocine and dihydrocodeine tartrate Arterial blood gases were Pao2 33 mm Hg, Paco, 56 mm (’DF118’) needed five or six maximal doses of one or other bicarbonate 25 mmol/1; pH 7.20. Electroencephalography Hg; the withdrawal episode fetal tachycardia with drug. During showed generalised slow waves but no focal or paroxysmal feapoor baseline variability and reduced transient changes associscan was normal; right carotid arteriogram E.M.I. tures; ated with fetal movements or Braxton-Hicks contractions were demonstrated a grossly hyperdynamic circulation; electroonly observed. Intramuscular DF118 was administered. A second and nerve-conduction studies were normal, as was myography taken 30 min later showed a to normal fetal return recording the cerebrospinal fluid. Thorough serology and culture failed heart-rate, but poor variability and diminished changes perto reveal a viral agent responsible for a recent encephalitic sisted. episode. Stresses which result in retarded in-utero growth are associAir encephalography 4 months later showed some dilatation ated with a reduced incidence of hyaline-membrane disease. 23 of the cerebral ventricles and a mild degree of cortical atrophy, The offspring of addicts may show evidence of growth disturShe 4 improved dramatically on mechanical ventilation, and bance.’ A lower incidence of hyaline-membrane disease has 7 months after admission she requires ventilation at midday been reported in neonates of animals5 and human mothers and at night when her sleeping respiration becomes inadeaddicted to heroin,67but the concept of fetal stress and materquate. Neurologically she remains normal, and in the daytime nal addiction remains a subject of deliberation.s Glass et a1.8 she is an active child learning new skills. Her only other probfound no difference in cord-blood cortico-steroid levels from lem is water retention if midday ventilation is insufficient. fetuses of addicted mothers and those of controls. Cord-steroid Some patients with Leigh’s subacute necrotising encephalobe influenced the stresses of values, however, may by delivery, myelopathy develop a primary (or central) hypoventilation and the incidence of hyaline-membrane disease need not reflect syndrome ("Ondine’s curse").2 Investigations in this girl have stresses inflicted on the fetus by drug withdrawal.9 revealed certain biochemical similarities to those reported in The above observations suggest the fetus may be subjected several cases of Leigh’s disease-namely, blood pyruvate 99 to "stress" in utero when maternal drug levels are withdrawn mol/1 (normal adult 35-60), blood lactate 3.06 mmol!1 rapidly. Falling maternal urinary oestrogen excretion often in(0-99-1-76), and a positive urine test for thiamine pyrophosdicates fetal compromise, and this falling trend is seen when adenosine triphosphate phosphoryltransferase inhibitor phate cut. It is also not uncommon for drug dosages are severely substance (Prof. J. R. Cooper).3 addicts to report increased fetal movements before their own At least two children with Ondine’s curse and a positive subjective symptoms of withdrawal. Cardiotocographic findurine test have been recorded. One died suddenly in hospital ings suggest evidence that there may also be associated fetal and the neuropathology showed hypothalamic necrosis, brainstress. stem inflammation, but not the typical features of Leigh’s disThe rapid withdrawal of drugs from addicted mothers may ease.4 Clearly a definitive diagnosis in this girl has not been have deleterious effects on their fetuses and should therefore reached especially if one considers Leigh’s disease to be a be avoided. Antenatal drug weaning programmes should be group of different enzyme or coenzyme disorders. This unfortunate child may be unique in presenting at thIs 1. Tylden, B. Adverse Drug Bull, 1973, 38, 120. age with central hypoventilation whilst asleep, yet remains 2. Gluck, L., Kulovich, M. V. Am. J. Obstet. Gynec. 1973, 115, 547. 3. Thomas, D. B. Aust. Pœdiat. J. 1975, 11, 26. developmentally normal and with no features of overt hypo4. Cochm, J. Fedn Proc. 1970, 20, 19. thalamic disease. The neuroradiological evidence of a braic toms
.
5. Taeusch, H. W. Jr., Carson, S. H., Wang, N. S., Avery, M. E. J. Pediat. 1973, 82, 869. 6 Sussman, S Amr. J. Dis. Child. 1963, 106, 325. 7. Glass, L., Rajegowda, B. K., Evans, H. G. Lancet, 1971, ii, 685. 8. Glass, L., Rajegowda, B. K., Mukherjee, T. K., Roth, M. M., Evans, H. G. Amr. J. Obstet. Gynec. 1973, 117, 416. 9. Zelson, C., Lee, S. J., Casalino, M. New Engl. J. Med. 1973, 289, 1216.
1. Severinghaus, J. W., Mitchell, R. A. Clin. Res. 1962, 10, 2. Pincus, J. H. Personal communication. 3. Cooper, J. R., Pincus, J. H., Itokawa, Y., Piros, K. New 4.
122.
Engl. JMed 1970, 283, 793. Pincus, J. H., Cooper, J. R., Piros, K., Turner, V. Neurology, 1974, 24, 885.
589
disorder suggests that the ultimate prognosis but further comments would be welcomed. Bristol Royal Hospital for Sick Children, Bristol BS2
8BJ
must
be
guarded
PETER G. F. SWIFT
ONE-HOUR BLOOD-XYLOSE TEST IN DIAGNOSIS OF COW’S-MILK ALLERGY
SIR,-Cow’s-milk allergy (C.M.A.) leads to a wide range of symptoms in infants and young children. At present, C.M.A. remains a clinical diagnosis based on a challenge procedure which should be repeated, ideally, at least three times: symptoms appear while the patient is on a milk-containing diet but disappear when milk is withdrawn.’ Serum-IgE level, milk antibodies, skin reactions, faecal fat, and xylosuria have been shown to be of limited help in the diagnosis of C.M.A.23 Complement activation4 and histoimmunological changes in jejunal mucosaJ have been reported after milk feeding in children with c.M.A. and are proposed as tests of diagnostic value. We wish to report preliminary results of the one-hour bloodxylose test in four children, 3 to 10 months old, who were challenged with cow’s milk between November, 1975 and March, 1976. They were suspected of having C.M.A. because of vomiting, chronic diarrhoea, failure to thrive, and microscopical detection of blood in stools. They all had a family history of allergic disorders. The 60 min blood-xylose tolerance was below 15 mg/dl in their initial diagnostic work-up while receiving cow’s milk. Our control value obtained in 50 normal children is 49+12-7 mg/dl after a load of 14.5 g of xylose 1m2 of body surface given in a 10% solution. All patients made good progress when changed to a milk-free diet contain-
ing gluten. After 4-12 wk the infants were readmitted for challenge with whole cow’s milk. A baseline 60 min blood-xylose test and jejunal biopsy for histological examination and disaccharidase activity were done before and 96 h after reintroduction of cow’s milk in the diet. Two of the four patients reacted clinically to the milk (see table). The histological appearances of the jejunal-biopsy specimens remained normal in two patients, whereas inflammatory cell infiltration was noticed in the postchallenge specimens of the other two patients. Disaccharidase activities, which were measured in three of the four children, fell in the post-challenge specimens. An important fall in the one-hour xylosaemia was observed in all four children 96 h after reintroduction of milk. These preliminary data suggest that the one-hour blood-xylose test could be valuable in the diagnosis of C.M.A. Department of Pediatrics, Division of Gastroenterology, Sainte-Justine Hospital, 3175 Sainte-Catherine Road,
Montreal, Quebec, H3T 1C5, Canada
CLAUDE L. MORIN
JEAN-PAUL BUTS ANDRÉE WEBER CLAUDE C. ROY
Goldman, A. S., Anderson, D. W., Sellers, W. A., Saperstein, S., Kniker, W. T, Halpern, S. R. Pediatrics, Springfield, 1963, 32, 425. 2. Freier, S. Clin. Allergy, 1973, 3, suppl. p. 597. 3 Kutitunen, P., Visakorpi, J. K., Savilahti, E., Pelkonen, P. Archs Dis. Childh. 1975, 50, 351. 4 Matthews, T. S., Soothill, J. F. Lancet, 1970, ii, 893. 5. Shiner, M., Ballard, J., Brook, C. G. D., Herman, S. ibid. 1975, ii, 1060.
ONE-HOUR XYLOSE TEST AND DISACCHARIDASE ACTIVITIES IN
c’
pressed as units per g of mucosal protein.
StR,—The possibility of interrupting pregnancy by giving antisera to hormones and placental antigens has long been of theoretical and practical interest.’ Loewit et al. have shown that this aim can be achieved with rabbit antisera to bovine luteinising hormone (bL.H.) injected into pregnant rats. In extending these investigations we have attempted to localise the antigen-antibody reaction in this experimental system. Rabbit antisera were prepared against bL.H. and rendered
specific by appropriate absorptions. Serological analyses were performed by double diffusion in gel and passive hæmagglutination, as well as indirect immunofluorescence (LF.) tests on frozen sections of rat pituitary glands. To check the in-vivo effect of the antiserum, pregnant rats received a single intraperitoneal injection between days 7 and 12 of pregnancy of either undiluted serum or of twofold serial dilutions up to 1/64. Normal rabbit serum (N.R.S.) and rabbit anti-normal bovine serum and anti-bovine prolactin served as controls. Day 11 seemed to be the most sensitive time for this kind of immunological intervention where dilutions of 1/64 of the antiserum were still effective in interrupting pregnancy. To demonstrate any damage to the kidneys in this process another group of pregnant and non-pregnant rats was treated with the same doses of antiserum in dilutions up to 1/8 or N.R.s., respectively. The animals were killed at various times after inoculation and the kidneys were snap frozen in liquid nitrogen for subsequent analyses in direct i.F. 4 µm frozen, unfixed sections were treated with a fluorescein-isothiocyanatelabelled swine anti-rabbit Ig serum. Bright linear and granular staining occurred along the glomerular basement membrane in rats treated with anti-bL.H., whereas the kidneys of the controls remained negative. To verify that the deposits of rabbit Ig were part of immune complexes with bL.H. the rabbit Ig was eluted for 20 min at room temperature from the kidney sections by means of a glycine hydrochloride buffer (pH 2-4). Sections so treated became in direct LF. with the antirabbit Ig conjugate. Reincubation of eluted sections with the anti-bL.H. serum, however, led to binding of specific antibody to the remaining L.H. not affected by the elution procedure, thus proving the specificity of the immune complexes. Prolonged elution (e.g., 60 min) led to the complete disappearance of both Ig and L.H.
negative
These results suggest that at least part of the antigen-antireaction in this experimental system takes place in the circulation and clearly show that immune-complex deposition does occur in the glomeruli of anti-bL.H.-treated rats. In view of the growing interest for immunological fertility control in humans this point might be of practical relevance.
body
1.
-
IMMUNE COMPLEXES IN KIDNEYS OF RATS TREATED WITH ANTISERUM TO LUTEINISING HORMONE
JEJUNAL
R. KOFLER G. WICK K. LOEWIT
Institute for General and
Experimental Pathology, Innsbruck, Austria
6020
E. (editor) Immunological Approaches Karolinska Institute, Stockholm, 1974. 2. Loewit, K., Badawy, S., Laurence, K. Endocrinology,
1 Diczfalusy,
MUCOSAL SPECIMENS BEFORE AND AFTER
COW’S
to
Fertility Control.
1969, 84, 244.
MILK PROVOCATION TEST
SiR,—There have been several reports of withdrawal symp-
well in advance to allow time for drawal of the drug of dependency.
planned
a
gradual with.
in neonates whose mothers were still addicted at delivery. Department of DAVID T. Y. LIU Obstetrics and Gynæcology, These babies appear normal at birth but within 10-36 h ELIZABETH TYLDEN University College Hospital become distressed and require treatment in a neonatal unit.’1 SINA H. TUKEL Medical School, London WC1. It is, therefore, in the infants’ interest that these addicted mothers should be identified and an attempt made to wean them off drugs before delivery. In the past five years 18 known addicts delivered in University College Hospital, London, and ONDINE’S CURSE 11 of these patients were weaned off opiates during pregnancy. readers SIR,Your may interested in a further example It is, however, not generally realised that the fetus may react Ondine’s curse in a young girl. The mythological reference of if or withdrawn adversely drugs are withheld during labour was first applied to three adults who had apnreic periods when rapidly during the antenatal period. Withholding drugs from awake but breathed on command and required mechanical known, or unsuspected, addicts has resulted in a stillbirth and ventilation when asleep.’Ondine, a German water nymph, ina neonatal death in neighbouring hospitals. voked a curse upon her jilted husband so that he would forget to the usually recogDuring labour, fetal distress-not due to breathe (and die) when he fell asleep. nised causes-arising in opiate addicts often appears to be The girl aged 2 years 8 months was admitted with a history relieved by the administration of pethidine or a replacement of recent mild diarrhoea and a family "flu-like" illness. For5 drug such as physeptone. This therapy is now advised in cases she had been intermittently drowsy and prone to sleep. days of opiate dependence. Immediately before admission she became difficult to rouse and We found that during the antenatal period, 24 h urinary cyanosed. During the preceding 9 months the parents had oestrogen production fell for the 48 h after each stepped reducnoticed that "she was blue around the lips" on three occasions tion in drug dosage. Thereafter, the level of maternal oestrogen in the early morning. She was otherwise healthy and had a excretion gradually rose until the drug was reduced again. On normal perinatal history. two occasions drug withdrawal had to be slowed because of Examination revealed a pale, cyanosed child with peripheral precipitate falls in urinary oestriols. limb oedema, puffy face, bounding pulses, and a soft precordial On one occasion, adverse changes in fetal heart-rate were murmur. The cyanosis was reversed by oxygen. Later she observed when external cardiotocography was carried out on became more drowsy with deep cyanosis, papilloedema, and a a patient during an episode of rapid drug withdrawal. A pacranial bruit. tient dependent on pentazocine and dihydrocodeine tartrate Arterial blood gases were Pao2 33 mm Hg, Paco, 56 mm (’DF118’) needed five or six maximal doses of one or other bicarbonate 25 mmol/1; pH 7.20. Electroencephalography Hg; the withdrawal episode fetal tachycardia with drug. During showed generalised slow waves but no focal or paroxysmal feapoor baseline variability and reduced transient changes associscan was normal; right carotid arteriogram E.M.I. tures; ated with fetal movements or Braxton-Hicks contractions were demonstrated a grossly hyperdynamic circulation; electroonly observed. Intramuscular DF118 was administered. A second and nerve-conduction studies were normal, as was myography taken 30 min later showed a to normal fetal return recording the cerebrospinal fluid. Thorough serology and culture failed heart-rate, but poor variability and diminished changes perto reveal a viral agent responsible for a recent encephalitic sisted. episode. Stresses which result in retarded in-utero growth are associAir encephalography 4 months later showed some dilatation ated with a reduced incidence of hyaline-membrane disease. 23 of the cerebral ventricles and a mild degree of cortical atrophy, The offspring of addicts may show evidence of growth disturShe 4 improved dramatically on mechanical ventilation, and bance.’ A lower incidence of hyaline-membrane disease has 7 months after admission she requires ventilation at midday been reported in neonates of animals5 and human mothers and at night when her sleeping respiration becomes inadeaddicted to heroin,67but the concept of fetal stress and materquate. Neurologically she remains normal, and in the daytime nal addiction remains a subject of deliberation.s Glass et a1.8 she is an active child learning new skills. Her only other probfound no difference in cord-blood cortico-steroid levels from lem is water retention if midday ventilation is insufficient. fetuses of addicted mothers and those of controls. Cord-steroid Some patients with Leigh’s subacute necrotising encephalobe influenced the stresses of values, however, may by delivery, myelopathy develop a primary (or central) hypoventilation and the incidence of hyaline-membrane disease need not reflect syndrome ("Ondine’s curse").2 Investigations in this girl have stresses inflicted on the fetus by drug withdrawal.9 revealed certain biochemical similarities to those reported in The above observations suggest the fetus may be subjected several cases of Leigh’s disease-namely, blood pyruvate 99 to "stress" in utero when maternal drug levels are withdrawn mol/1 (normal adult 35-60), blood lactate 3.06 mmol!1 rapidly. Falling maternal urinary oestrogen excretion often in(0-99-1-76), and a positive urine test for thiamine pyrophosdicates fetal compromise, and this falling trend is seen when adenosine triphosphate phosphoryltransferase inhibitor phate cut. It is also not uncommon for drug dosages are severely substance (Prof. J. R. Cooper).3 addicts to report increased fetal movements before their own At least two children with Ondine’s curse and a positive subjective symptoms of withdrawal. Cardiotocographic findurine test have been recorded. One died suddenly in hospital ings suggest evidence that there may also be associated fetal and the neuropathology showed hypothalamic necrosis, brainstress. stem inflammation, but not the typical features of Leigh’s disThe rapid withdrawal of drugs from addicted mothers may ease.4 Clearly a definitive diagnosis in this girl has not been have deleterious effects on their fetuses and should therefore reached especially if one considers Leigh’s disease to be a be avoided. Antenatal drug weaning programmes should be group of different enzyme or coenzyme disorders. This unfortunate child may be unique in presenting at thIs 1. Tylden, B. Adverse Drug Bull, 1973, 38, 120. age with central hypoventilation whilst asleep, yet remains 2. Gluck, L., Kulovich, M. V. Am. J. Obstet. Gynec. 1973, 115, 547. 3. Thomas, D. B. Aust. Pœdiat. J. 1975, 11, 26. developmentally normal and with no features of overt hypo4. Cochm, J. Fedn Proc. 1970, 20, 19. thalamic disease. The neuroradiological evidence of a braic toms
.
5. Taeusch, H. W. Jr., Carson, S. H., Wang, N. S., Avery, M. E. J. Pediat. 1973, 82, 869. 6 Sussman, S Amr. J. Dis. Child. 1963, 106, 325. 7. Glass, L., Rajegowda, B. K., Evans, H. G. Lancet, 1971, ii, 685. 8. Glass, L., Rajegowda, B. K., Mukherjee, T. K., Roth, M. M., Evans, H. G. Amr. J. Obstet. Gynec. 1973, 117, 416. 9. Zelson, C., Lee, S. J., Casalino, M. New Engl. J. Med. 1973, 289, 1216.
1. Severinghaus, J. W., Mitchell, R. A. Clin. Res. 1962, 10, 2. Pincus, J. H. Personal communication. 3. Cooper, J. R., Pincus, J. H., Itokawa, Y., Piros, K. New 4.
122.
Engl. JMed 1970, 283, 793. Pincus, J. H., Cooper, J. R., Piros, K., Turner, V. Neurology, 1974, 24, 885.
589
disorder suggests that the ultimate prognosis but further comments would be welcomed. Bristol Royal Hospital for Sick Children, Bristol BS2
8BJ
must
be
guarded
PETER G. F. SWIFT
ONE-HOUR BLOOD-XYLOSE TEST IN DIAGNOSIS OF COW’S-MILK ALLERGY
SIR,-Cow’s-milk allergy (C.M.A.) leads to a wide range of symptoms in infants and young children. At present, C.M.A. remains a clinical diagnosis based on a challenge procedure which should be repeated, ideally, at least three times: symptoms appear while the patient is on a milk-containing diet but disappear when milk is withdrawn.’ Serum-IgE level, milk antibodies, skin reactions, faecal fat, and xylosuria have been shown to be of limited help in the diagnosis of C.M.A.23 Complement activation4 and histoimmunological changes in jejunal mucosaJ have been reported after milk feeding in children with c.M.A. and are proposed as tests of diagnostic value. We wish to report preliminary results of the one-hour bloodxylose test in four children, 3 to 10 months old, who were challenged with cow’s milk between November, 1975 and March, 1976. They were suspected of having C.M.A. because of vomiting, chronic diarrhoea, failure to thrive, and microscopical detection of blood in stools. They all had a family history of allergic disorders. The 60 min blood-xylose tolerance was below 15 mg/dl in their initial diagnostic work-up while receiving cow’s milk. Our control value obtained in 50 normal children is 49+12-7 mg/dl after a load of 14.5 g of xylose 1m2 of body surface given in a 10% solution. All patients made good progress when changed to a milk-free diet contain-
ing gluten. After 4-12 wk the infants were readmitted for challenge with whole cow’s milk. A baseline 60 min blood-xylose test and jejunal biopsy for histological examination and disaccharidase activity were done before and 96 h after reintroduction of cow’s milk in the diet. Two of the four patients reacted clinically to the milk (see table). The histological appearances of the jejunal-biopsy specimens remained normal in two patients, whereas inflammatory cell infiltration was noticed in the postchallenge specimens of the other two patients. Disaccharidase activities, which were measured in three of the four children, fell in the post-challenge specimens. An important fall in the one-hour xylosaemia was observed in all four children 96 h after reintroduction of milk. These preliminary data suggest that the one-hour blood-xylose test could be valuable in the diagnosis of C.M.A. Department of Pediatrics, Division of Gastroenterology, Sainte-Justine Hospital, 3175 Sainte-Catherine Road,
Montreal, Quebec, H3T 1C5, Canada
CLAUDE L. MORIN
JEAN-PAUL BUTS ANDRÉE WEBER CLAUDE C. ROY
Goldman, A. S., Anderson, D. W., Sellers, W. A., Saperstein, S., Kniker, W. T, Halpern, S. R. Pediatrics, Springfield, 1963, 32, 425. 2. Freier, S. Clin. Allergy, 1973, 3, suppl. p. 597. 3 Kutitunen, P., Visakorpi, J. K., Savilahti, E., Pelkonen, P. Archs Dis. Childh. 1975, 50, 351. 4 Matthews, T. S., Soothill, J. F. Lancet, 1970, ii, 893. 5. Shiner, M., Ballard, J., Brook, C. G. D., Herman, S. ibid. 1975, ii, 1060.
ONE-HOUR XYLOSE TEST AND DISACCHARIDASE ACTIVITIES IN
c’
pressed as units per g of mucosal protein.
StR,—The possibility of interrupting pregnancy by giving antisera to hormones and placental antigens has long been of theoretical and practical interest.’ Loewit et al. have shown that this aim can be achieved with rabbit antisera to bovine luteinising hormone (bL.H.) injected into pregnant rats. In extending these investigations we have attempted to localise the antigen-antibody reaction in this experimental system. Rabbit antisera were prepared against bL.H. and rendered
specific by appropriate absorptions. Serological analyses were performed by double diffusion in gel and passive hæmagglutination, as well as indirect immunofluorescence (LF.) tests on frozen sections of rat pituitary glands. To check the in-vivo effect of the antiserum, pregnant rats received a single intraperitoneal injection between days 7 and 12 of pregnancy of either undiluted serum or of twofold serial dilutions up to 1/64. Normal rabbit serum (N.R.S.) and rabbit anti-normal bovine serum and anti-bovine prolactin served as controls. Day 11 seemed to be the most sensitive time for this kind of immunological intervention where dilutions of 1/64 of the antiserum were still effective in interrupting pregnancy. To demonstrate any damage to the kidneys in this process another group of pregnant and non-pregnant rats was treated with the same doses of antiserum in dilutions up to 1/8 or N.R.s., respectively. The animals were killed at various times after inoculation and the kidneys were snap frozen in liquid nitrogen for subsequent analyses in direct i.F. 4 µm frozen, unfixed sections were treated with a fluorescein-isothiocyanatelabelled swine anti-rabbit Ig serum. Bright linear and granular staining occurred along the glomerular basement membrane in rats treated with anti-bL.H., whereas the kidneys of the controls remained negative. To verify that the deposits of rabbit Ig were part of immune complexes with bL.H. the rabbit Ig was eluted for 20 min at room temperature from the kidney sections by means of a glycine hydrochloride buffer (pH 2-4). Sections so treated became in direct LF. with the antirabbit Ig conjugate. Reincubation of eluted sections with the anti-bL.H. serum, however, led to binding of specific antibody to the remaining L.H. not affected by the elution procedure, thus proving the specificity of the immune complexes. Prolonged elution (e.g., 60 min) led to the complete disappearance of both Ig and L.H.
negative
These results suggest that at least part of the antigen-antireaction in this experimental system takes place in the circulation and clearly show that immune-complex deposition does occur in the glomeruli of anti-bL.H.-treated rats. In view of the growing interest for immunological fertility control in humans this point might be of practical relevance.
body
1.
-
IMMUNE COMPLEXES IN KIDNEYS OF RATS TREATED WITH ANTISERUM TO LUTEINISING HORMONE
JEJUNAL
R. KOFLER G. WICK K. LOEWIT
Institute for General and
Experimental Pathology, Innsbruck, Austria
6020
E. (editor) Immunological Approaches Karolinska Institute, Stockholm, 1974. 2. Loewit, K., Badawy, S., Laurence, K. Endocrinology,
1 Diczfalusy,
MUCOSAL SPECIMENS BEFORE AND AFTER
COW’S
to
Fertility Control.
1969, 84, 244.
MILK PROVOCATION TEST
