NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
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RECEPTORS.
These according to Ehrlich's theory are side-chains in the central protoplasmic
m o l e c u l e o f a c e l l , c a p a b l e o f c o m b i n i n g w i t h o t h e r f r e e o r e x t e r n a l g r o u p s o r
receptors.; It is upon these combining affinities that the cell is dependent for its
normal metabolism ; but their existence also exposes it to the toxic effects we are
discussing. The receptors of one species of animal are foreign, matter in, the body
of anot her speci es ; but pr ovi ded t he cel l s of t he ani mal i nt o whi ch f or ei gn
blood has been injected possess side-chains or receptors capable of combining
with those of the injected blood cells, combination at once takes place. But when
the combination between the cell receptors of the injected animal and those of the
foreign blood cells has taken place, the former cells are stimulated to produce new
receptors to take the place of those already occupied.
These receptors are generally produced in excess and may be thrown, off becom-
ing free in, the blood, and thus giving rise to an anti-body, specific for cells similar
to those that stimulated its production. This anti-body possesses the power, of
combining with such foreign, cells both in vitro and when, injected in vivo ; and
if the necessary complement is present their combination will result in hæmolysis.
It is equally evident that upon the presence of suitable receptors wilt depend
the effect of the injection of such specific toxins as the hæmolysis. We have
seen that the injection into an. animal of foreign .blood cells, with their recep-
tors, will cause the cells of the animal injected to produce and throw off an excess
of receptors capable of combining with cells similar to these injected. These
free receptors are amboceptors specific for similar foreign, bloods, which necessa-
rily possess receptors with which they can combine. If then such free amboceptors
be injected in. turn into an animal 'whose blood cells possess the suitable recep-
tors, combination will ensue under certain, conditions we shall further indicate.
If on the contrary the blood cells of an animal into which such an, amboceptor
is injected, possess. no receptors with which it can, combine, combination cannot
occur and no reaction, will ensue.
It mill be necessary to refer more fully to the subject of receptors later on.
AMBOCEPTORS.
These, as we have already seen, are the free cell receptors that have been
cast off in response to stimulation, by foreign receptors. They possess in general
certain properties whichit is necessary to study—
(1) The power of combining with cells by means of a cytophile affinity
.
(2) The power of combining with complement by means of their complemento-
phile affinity.
I P / Q B , 1 0
No. 16.
(NEW SERIES. )
SCIENTIFIC MEMOIRS
BY
OFFICERS OF THE MEDICAL AND SANITARY DEPARTMENTS
OF THE
GOVERNMENT OF INDIA.
THE SPECIFICITY OF ANTIVENOMOUS SERA WITH SPECIAL REFERENCE
TO A SERUM PREPARED WITH THE VENOM OF DABOIA RUSSELLI.
BY
CAPTAIN GEORGE LAMB, M.D., I.M.S.
ISSUED UNDER THE AUTHORITY OF THE GOVERNMENT OF INDIA
BY THE SANITARY COMMISSIONER WITH THE GOVERNMENT
OF INDIA, SIMLA.
CALCUTTA :
OFFICE OF THE SUPERINTENDENT OF GOVERNMENT PRINTING, INDIA.
1905.
Price Annas 6 or 7d.
No. 16.
(NEW SERIES.)
SCIENTIFIC MEMOIRS
BY
OFFICERS OF THE MEDICAL AND SANITARY DEPARTMENTS
OF THE
GOVERNMENT OF INDIA.
THE SPECIFICITY OF ANTIVENOMOUS SERA, WITH SPECIAL REFERENCE
TO A SERUM PREPARED WITH THE VENOM OF DABOIA RUSSELLII.
BY
CAPTAIN GEORGE LAMB, M.D., I.M.S.
ISSUED UNDER THE AUTHORITY OF THE GOVERNMENT OF INDIA
BY THE SANITARY COMMISSIONER WITH THE GOVERNMENT
OF INDIA, SIMLA.
CALCUTTA:
OFFICE OF THE SUPERINTENDENT OF GOVERNMENT PRINTING, INDIA.
1905.
Agents for the Sale of Books published by the Superintendent of Government
Printing, India, Calcutta.
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ON THE CONTINENT.
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Press, Lahore.
OTTO HARRASSOWITZ, Leipzig.
Superintendent, American Baptist Mission Press,
Rangoon.
KARL W. HIERSEMANN, Leipzig.
THE SPECIFICITY OF ANTIVENOMOUS SERA, WITH SPECIAL REFERENCE
TO A SERUM PREPARED WITH THE VENOM OF DABOIA RUSSELLII.
IN two previous communications1 on the specificity of antivenomous sera I
brought forward evidence which pointed to the conclusion that a pure
antivenomous serum, that is, a serum prepared with a single poison, is markedly,
if not absolutely, specific for the venom which has been used in its preparation.
This conclusion was based on the results of the testing of two antivenomous
sera, one prepared with pure cobra poison and the other with the pure poison of
the Australian tiger snake (Hoplocephalus curtus), against the general actions
in vivo, the hæmolytic actions in vitro and the actions on the coagulability
of the blood plasma also in vitro, of ten different venoms, some of which were
the poisons of colubrine species while others were of viperine origin. It is un-
necessary to enter into the details of the experiments which were then carried out
and which have been set forth in the papers above mentioned. Before, however,
passing on to the subject matter proper of the present communication, to wit,
some observations made with the object of the testing of a serum prepared with a
pure viperine venom, that of Daboia Russellii, I have to refer briefly to the results
which were obtained in vivo with the cobra venom anti-serum and the poisons
of some of the other colubrine snakes against which it was tested. This refer-
ence is necessary in view of the fact, that the results which have been obtained by
Captain Rogers2with Calmette's serum differ from those which I have obtained
with the serum prepared with pure cobra venom. It is, however, to be remem-
bered in this connection that Calmette's serum is made by vaccinating with a
mixture of venoms, of which mixture cobra venom is the preponderating consti-
tuent. This fact alone might have been brought forward as an adequate
explanation of the differences between my results and those obtained by Captain
Rogers, had not this observer in a more recent paper 3 stated that, using a
serum prepared in London by Messrs. Burroughs and Wellcome with pure cobra
venom, he obtained similar results to those which he had previously got with
Calmette's serum. Further, in a letter to the 'Lancet' Captain Rogers 4 has
recently stated, that Dr. Calmette has told him that the serum with which he
was supplied and which he used for his experiments happened to be prepared
with pure cobra venom. His experiments, therefore, are strictly comparable
with those which I have made with a pure cobra venom anti-serum. In what
then do the differences consist, and how are they to be explained?
B
2
In the first place, let us consider the experiments with the venom of the
king cobra, a snake belonging to the same genus as the cobra. Captain Rogers,
working with a serum, 1 c.c. of which, he showed, was able to neutralise 0.56 milli-
gramme of cobra venom, found that the same quantity of serum could completely
neutralise 0.3475 milligramme of the venom of the king cobra, the toxicity of
which poison is practically the same as that of cobra venom. Now, in my ex-
periments with a cobra venom anti-serum, 1 c.c. of which was able to neutralise
1.5 milligrammes of its homologous poison, it was found that, while large
amounts of serum were able to delay death in cases of intoxication with the
venom of the king cobra, no complete neutralisation took place : that 1 c.c. of
serum could not neutralise even 0.13 milligramme of this poison. It will be seen,
therefore, that while the serum I used was, as far as cobra venom was concerned,
nearly three times as strong as that employed by Captain Rogers, it failed to
neutralise about a third of the amount of king cobra venom which this observer
found Calmette's serum could effectually neutralise.
It is to be noted that in both series of experiments the sera were tested
against ten lethal doses of venom, but that while Captain Rogers used pigeons I
employed rabbits. I have recently repeated the observations which bear on this
point. In these experiments a serum prepared with pure unheated cobra venom
was again employed : 1c.c. of this serum was able to neutralise 0.7 milligramme
of pure cobra venom. It was now tested against only 6 lethal doses of king cobra
venom by subcutaneous injection into rabbits, the serum and venom being
allowed to stand in vitro at room temperature for half an hour before injection.
A reference to the protocols (table I) will show that even 15 c.c. of serum
could not save the life of the animal. We can, moreover, calculate that 1 c.c.
of serum had failed to neutralise 0.11 milligramme of this poison. From this
series of experiments, therefore, we arrive at the same conclusion as we had
come to previously, namely, that, while a serum prepared with pure cobra venom
and strongly antitoxic for this venom delays death in cases of intoxication with
the venom of the king cobra, it does not completely neutralise this poison even
when used in large quantities.
It is difficult to reconcile my results with those obtained by Captain Rogers.
It is, however, to be noted that Captain Rogers' experiments were made with
pigeons while mine were carried out with rabbits. It might be conceivable that a
certain action of the poison in mammals was wanting in birds, and that the con-
stituent which caused this action remained unneutralised in my experiments.
That all the constituents of king cobra venom are not neutralised by a cobra
antivenom is evident, as I have shown,5from the fact that such a serum has no
hindering effect on the hæmolytic action of king cobra poison when tested
in vitro. Further, Rogers does not definitely state that in his experiments the
3
minimum lethal dose of the venom was determined for pigeons, the animals on
which the serum experiments were carried out : in fact, by stating that, on account
of a difficulty in obtaining rats, pigeons were used for the serum experiments, he
leads one to suppose that the minimum lethal dose had been determined for rats
and not for pigeons. It is possible, therefore, that in his serum experiments with
pigeons he was using not ten lethal doses but a very much smaller number. The
only other possible explanation that I can think of is that the poison with which
Captain Rogers was working was not that of the king cobra. I may state that the
king cobra venom which I used was collected directly under my own supervision
or that of Colonel Bannerman, I.M.S., from snakes the property of the
Natural History Society, Bombay.
In the second place we have to consider the experiments with the venom
of Bungarus cœruleus or common krait.
Working with Calmette's serum Captain Rogers found that 1 c.c. could
neutralise 0.072 milligramme of krait poison. In my experiments it was found
that 1 c.c. of pure cobra venom anti-serum, which was nearly three times
the strength of the serum used by Captain Rogers, failed to neutralise 0.0925
milligramme of the venom of Bungarus cœruleus and that, moreover, the serum
had no effect on the progress of the intoxication.
I have recently repeated these experiments, again using a pure cobra anti-
v e n o m , 1 c . c . o f w h i c h w a s a b l e t o n e u t r a l i s e 0 . 7 m i l l i g r a m m e o f c o b r a p o i s o n .
Employing in these experiments as a test dose only 5 lethal doses by intra-
venous injection into rabbits, I found that 8 c.c. of serum had no effect whatever
in even prolonging life (protocols, table II). We can, therefore, calculate that
1 c . c . o f t h i s s e r u m f a i l e d t o n e u t r a l i s e 0 . 0 2 5 m i l l i g r a m m e o f v e n o m , a r e s u l t
strictly comparable to that previously arrived at.
A careful perusal of Captain Rogers' paper shows that a serious objection
can be at once raised to his experiments. For, it is seen that, while he determined
the minimum lethal dose for pigeons, he used rats in the serum experiments,
evidently presuming the venom to be of the same toxicity for these latter animals
as for pigeons. Thus, he found the minimum lethal dose for pigeons to be 0.25
milligramme per kilo., and accordingly in the serum experiments he used as a test
dose 2.5 milligrammes per kilo. for rats. Now, in a more recent paper 6 from Pro-
fessor Fraser's laboratory, Elliot, Sillar and Carmichael have accurately determined
the minimum lethal dose of this venom for rats and have found it to be one milli-
gramme per kilo. It is, therefore, apparent that in his serum experiments Captain
Rogers was using as a test dose only 2½ lethal doses instead of ten lethal
doses as he supposed. Further, it is doubtful whether he was working with pure
krait venom. For, as a result of his experiments on the physiological actions he
states, that the symptoms following poisoning by this venom are identical with
B 2
4
those following poisoning by cobra venom, whereas the results obtained by the
three workers mentioned above indicate, " that while the symptoms are similar,
still they differ so much in relative degree as to render it doubtful if they can in
future be spoken of as identical." Further, I hope to show at a later date that
the action of this venom on the central nervous system is quite different from that
of the poison of the cobra. I may state that the krait venom which I used for
my observations was collected at the Plague Research Laboratory, Bombay,
either under my own supervision or that of Colonel Bannerman.
As regards the experiments with the venoms of the other colubrine snakes
Captain Rogers and I are in agreement, namely, that in relatively large amount
a cobra venom anti-serum is able to neutralise in vivo the poison of one of the sea
snakes, Enhydrina vaiakadien, while it delays death but does not neutralise
the venom of Bungarus fasciatus. In this latter case Captain Rogers is of opinion
that the colubrine constituents of the venom are neutralised, the animal dying of
chronic viperine poisoning. To this point I shall again refer in a paper, to be
written along with Dr. Hunter, on the action of viperine poisons on the central
nervous system.
From these considerations it seems to me that the conclusions arrived at
in my last paper on the specificity of antivenomous sera were quite justified,
namely, that a serum of a horse immunised with pure cobra poison is strongly
antitoxic for the venom used in its preparation : when used in large quantity it
has a slight neutralising power for the venom of Enhydrina valakadien, one of the
common sea snakes : further, it delays death in cases of intoxication with the
venom of the king cobra, a species belonging to the same genus as the cobra,
and also in cases of intoxication with the venom of Bungarus fasciatus : it does
not, however, completely neutralise these poisons even when used in large
quantities. The serum would be of little or no therapeutic value in cases of
bites from these three snakes. Finally, the serum contains no antitoxic sub-
stances, which are active against the venom of Bungarus cœruleus. It is to
be remembered that these conclusions refer only to the results of the testing
of the serum ' i n v i v o ' .
We are not concerned at present with the hæmoly-
tic actions or the actions on the blood plasma of the several venoms as tested
in vitro.
We may now pass on to the consideration of the subject matter
proper of the present communication, that is, some observations which were
made with an anti-serum for the venom of Daboia Russellii. This serum
was prepared by injecting a horse with gradually increasing doses of the
pure unheated poison of Russell's viper. The treatment lasted over a period of
one year. From a dose of one milligramme the amount given at each injec-
tion was gradually increased until at the time of the testing of the serum
5
a dose of 500 milligrammes could be given without causing any disturbance other
than considerable local reaction. The serum was now tested against the same ten
poisons as were used in the testing of the cobra venom anti-serum. Further, the
same plan of testing was adopted in this case as was employed on the previous
occasion. Thus, in the first place, the serum was tested against the general
actions in vivo of the several poisons : in the second place, it was tested against
their hæmolytic actions in vitro and, in the third place, it was tested against the
actions of several of the poisons on the coagulability of the blood plasma
also in vitro. It will be seen, therefore, that in this case also the serum has been
tested in a fairly exhaustive manner.
Let us first consider the experiments made with the object of ascertaining
if the serum had any neutralising effect on the general actions in vivo of the
various poisons.
Colubrine snakes. —The serum was tested against the venoms of five colu-
brine snakes, namely, cobra, king cobra, Bungarus cœruleus, Bungarus fas-
ciatus and Enhydrina valakadien (Sea-snake). The results of these obser-
vations are detailed in the protocols (table III). From these it will be seen that,
as was to be expected, although comparatively large amounts of serum were used,
no neutralising action was exhibited by the serum in any instance.
Viperine snakes.—The poisons of four viperine species were available for
testing the serum. These were, (1) Daboia Russellii, (2) Echis carinata ( phoorsa),
(3) Trimeresurus gramineus (Green pit viper) and (4) Crotalus adamanteus (the
American rattlesnake).
Daboia Russellii. —A series of careful experiments was made with this poison
in order to standardise accurately the serum against the venom with which it
was prepared. The test dose used was fifty lethal doses by intravenous injection
into rabbits, namely, 5 milligrammes per kilo. A reference to the protocols (table
IV) will show, that 1.75 c.c. of serum were able to save the life of the animal
when mixed with this test dose. Thus, it can be calculated that 1 c.c. of serum
would be able to neutralise 2.8 milligrammes of venom. Further, it will also be seen
that, while the animal which received 0.75 c.c. of serum along with the test dose
of venom died in two minutes, the result of intravascular clotting, the rabbit which
got 1 c.c. of serum lived for 10½ hours. In this latter case the serum had evidently
neutralised the fibrin ferment of the poison, but had failed to neutralise the consti-
tuents which cause the more or less chronic effects. In a previous communication 7
I have stated that the strongest cobra venom anti-serum which I have been able to
prepare was one of which 1 c.c. could neutralise 1.5 milligrammes of pure cobra
venom, but that as a rule it was only possible to obtain a serum of much less
potency than this. It would, therefore, appear that daboia venom yields a stronger
serum than cobra venom. The horse from which the present serum was obtained
6
had been under treatment for only a year, and the maximum amount of venom it
had received at one injection was 500 milligrammes.
Echis carinata .—The physiological action of this venom, as far as I have
ascertained from a number of experiments, is similar to that of daboia venom.
It is, however, of somewhat greater toxicity. It was, therefore, of the greatest
interest to ascertain if daboia venom anti-serum had any neutralising effect on
the general action in vivo of this poison.
In a preliminary series of experiments a test dose of ten lethal doses, namely,
0.5 milligramme per kilo. by intravenous injection into rabbits, was used. As, in
these experiments, the serum appeared to have no effect whatever on the action of
the poison, a second observation was made in which the test dose used was reduced
to two lethal doses. A reference to the protocols, table V, will show, that an
animal which received 5 c.c. of serum along with this test dose died in the same
time as the control. We can, therefore, calculate that 1 c.c. of serum could not
neutralise the general action in vivo of 0.012 milligramme of this poison, a quantity
more than two hundred times less than the amount of daboia venom which 1 c.c.
of the same serum could neutralise. Further, an experiment was made in which
a single lethal dose of venom was used : 5 c.c. of serum in this case had no effect
in preventing or even delaying death, which took place 5 minutes after the injection.
It is evident, therefore, that this serum has no effect on the general action
in vivo of the venom of Echis carinata. This conclusion is of considerable interest
in view of the fact, to be referred to immediately, that the serum has a strong
neutralising effect on the hæmolytic action of this poison when tested in vitro .
Trimeresurus gramineus. —This snake is a small Indian viper of the
family Crotalinæ, the same family to which the American rattlesnake belongs.
Unfortunately I have not been able to obtain sufficient poison to enable me to
work out its physiological action in any detail. From a few experiments which
have been made it can be said that the action of the poison is similar, if not
identical, to that of the venom of Crotalus adamanteus : it is not, however, nearly
so toxic as this latter poison.
The test dose used to ascertain if daboia venom anti-serum has any neu-
tralising effect on the general action in vivo of this poison was 2 milligrammes
per kilo. by intravenous injection into rabbits, an amount less than two lethal doses.
It was found that 5 c.c. of serum were unable to prevent death in such an experi-
ment (protocols, table VI), We may conclude, therefore, that the serum has
no neutralising effect on the action in vivo of this poison.
Crotalus adamanteus. —The minimum lethal dose of this venom for a rabbit
by intravenous injection was found to be 0.25 milligramme per kilo. The test dose
now used to test the serum under experiment was 8 lethal doses, that is to say, 2
milligrammes per kilo. by intravenous injection. It was found that 4 c.c. of serum