Part 13

We must emphasise the fact, established by later researches, that after extirpation of the spleen, an enlargement of various lymphatic glands occurs. The alterations of the thyroid, which have been observed by many authors, cannot be described as constant.

Further, the blood investigations which Mosler, Robin, Winogradow, Zersas and others have carried on in animals and man after removal of the spleen must here be mentioned. These have already proved that a leucocytosis occurs after some considerable time. Prof. Kurloff carried out detailed investigations in 1888 in Ehrlich's laboratory, and carefully studied the condition of the blood after extirpation of the spleen. As the work of Prof. Kurloff has so far only appeared in Russian, his important results may be here recorded more fully. For his researches, Kurloff employed the guinea-pig, as this animal by its peculiar blood is specially suited for this purpose.

In order to give a systematic account of the results of these important investigations, we must first shortly sketch the normal histology of the blood of the guinea-pig according to Kurloff.

In the blood of the healthy guinea-pig the following elements are found.

I. Cells bearing granules.

1. =Polynuclear, with pseudoeosinophil granulation.= This granulation, which Ehrlich had previously found in the rabbit, is easily distinguishable from the true eosinophil, since it is much finer, and stains quite differently in eosine-aurantia-nigrosin mixtures. One principal distinction between these two forms of cells lies in the fact that, according to Kurloff, this granulation is very easily dissolved by acid, but remains unchanged in alkaline solutions; doubtless an indication that the granulation consists of a basic body soluble with difficulty, which with acids forms soluble salts. The true eosinophil granulation remains, on the other hand, quite unchanged under these conditions.

=These pseudoeosinophil, polynuclear cells, correspond functionally to the neutrophil polynuclear of man=; their number amounts to 40-50% of the total white cells. The red bone-marrow is to be regarded as the place of origin of this kind of cell. It contains very many pseudoeosinophil cells, and indeed all stages are to be found in it, from the mononuclear cells bearing granules to the fully formed polynuclear.

2. The typical =eosinophil leucocytes=, which fully correspond to those found in man, and amount to about 10% of the number of the white.

3. The "=nigrosinophil cells=," as they are called by Kurloff. In their general appearance, in the size of the cell and the granulation, they completely correspond to the eosinophil cell. The only distinction between them consists in a chemical difference in the granulation. These cells stain in the colour of nigrosin in the aurantia-eosin-nigrosin mixture, whilst the eosinophil cells become red. The two granulations always show different shades in the triacid preparation as well; for the nigrosinophil cells stain a blacker hue.

II. Cells free from granules.

([alpha]) Cells with vacuoles.

This is a quite peculiar group, characteristic for the blood of the guinea-pig. It shews transitions in the blood, from large mononuclear to transitional and polynuclear forms, but is marked by the lack of any kind of granulation. Instead of the latter, we find in these cells a roundish, nucleus-like form in the protoplasm, which also takes the nuclear stains, and possibly is to be considered an accessory nucleus. We have received the impression that we have here to deal with a vacuole filled with substance secreted by the cell. In a large series of preparations, it is possible to obtain some elucidation of the development and fate of these appearances. They first appear as point-like granules in the protoplasm, bearing no relation to the cell nucleus; they gradually increase, and acquire a considerable circumference. When they have attained about the size of the cell nucleus, they, or rather their contents, appear to break through the protoplasmic membrane and to leave the cell.

The number of the vacuole containing cells is 15-20% of the colourless blood corpuscles.

() Typical lymphocytes.

Their appearance completely corresponds with that of human lymphocytes as described above. They make up 30-35% of the total number of leucocytes.

Now Kurloff in the course of extremely careful and laborious researches, estimated the total number of leucocytes, and then from the percentage numbers, the total quantity of pseudoeosinophil, neutrophil, eosinophil, vacuole containing cells, and lymphocytes, and could thus demonstrate that in uncomplicated cases of removal of the spleen, where inflammatory processes, accompanied by an increase of the polynuclear neutrophil corpuscles, were avoided, a =gradual increase of the lymphocytes= alone in course of time results. This may be a two- or threefold increase, whereas the numbers of all other elements remain unchanged.

Kurloff obtained his figures as follows: first he estimated the relative proportion of the different kinds of white blood corpuscles one to another in a large number of cells (500 to 1000). A count of this kind however gives no evidence as to whether one or other kind of cell is absolutely increased or diminished. A fall in the percentage of the lymph cells may be brought about by two quite different factors: (1) by a diminished production of lymphocytes, (2) by an increased influx of polynuclear forms, which naturally lowers the relative count of the lymphocytes. It was therefore necessary to obtain a method which would show alterations in the absolute number of the individual forms of leucocytes. Kurloff used for this purpose the "comparative field"; that is, he counted by the aid of a moveable stage the different forms which lay on a definite area (22 sq. mm.) of the dried blood preparation. This procedure gave very exact results, as only faultlessly prepared, and regularly spread preparations were used. The following figures (from Exp. II.) illustrate the method and its results:

April 12 52% pseudo-eos. 10% lymphocytes counted. Sept. 2 (one month after the operation) 22% " 53% " "

By the aid of the comparative surface, these figures were supplemented by the following averages. On each surface used for comparison were found:

April 12 38 white = 19.8 pseudo-eos. 10.6 lymphocytes. Sept. 2 81 " 18.0 " 46.9 "

From this example it follows without doubt, that the =total number of the white blood corpuscles had about doubled itself=, but that in this increase the lymphocytes exclusively were concerned, and the pseudo-eosinophil cells had not undergone the smallest increase.

The results which Kurloff obtained by means of this method in animals whose spleens had been removed, may be illustrated by one of his original researches and its accompanying chart and table.

Exp. I. Young female, weight 234 gr. Number of red corpuscles in a cubic millimeter of blood 5,780,000. Number of white 10,700. On April 19, 1888, the spleen was removed, the wound healed by first intention. The results of the further investigation of the blood are found in the following table.

From the chart and table, the number on the surface of comparison of the white blood corpuscles is seen to have more than doubled itself in the first seven months, and that this increase was solely dependent on the flooding of the blood by lymphocytes. The nucleated or bone-marrow elements and the large mononuclear cells remained continuously at the same level during the whole period. The changes in the percentage proportions ran somewhat differently. The percentages rose from 35 to 66% for the lymphocytes only, whilst for the other forms they distinctly fell: for the nucleated from 44% to 22% and for the large mononuclear from 18% to 9%. It was only in the course of the second year that a very considerable relative and absolute increase of the eosinophil cells appeared: the values rose gradually from about 1.0% to 28.9% or from 0.5 to 13.9 on each comparison area. The last examination of the blood in this animal was made on April 30, 1890, that is, two years after the removal of the spleen. The animal was quite healthy, bore four healthy young guinea-pigs by a father whose spleen had been removed. The young have a completely normal spleen, and their blood likewise shows no abnormalities.

Thick line--total number of leucocytes Broken line--lymphocytes Thin line--number of nucleated, pseudo-eosinophil cells Double line--large mononuclear cells Dotted line--eosinophil cells]

TABLE I.

Key to columns: A - Leucocytes B - Pseudo-eosinophil cells C - Lymphocytes D - Large mononuclear cells E - Eosinophil cells F - Nigrosinopil cells G - On comparative surfaces

--------------------------------------------------------------------------- || A || B || C || D || E || F | ------------------------------------------------------------------ Date ||Total| G|| % | G || % | G || % | G || % | G || % | G | --------------------------------------------------------------------------- 1888 || | || | || | || | || | || | | April 19|| 500|--||44.7| -- ||35.4| -- ||18.4| -- || 1.1| -- || 0.5| -- | 23|| 990|24||40.4| 9.7||35.6| 8.5||21.6| 5.2|| 1.9| 0.4 || 0.4|0.09| May 1|| 858|28||47.0|13.6||32.6| 9.1||18.0| 5.0|| 0.9| 0.2 || 0.3|0.08| 8|| 934|28||45.2|12.6||40.3|11.3||14.3| 4.0|| 0.6| 0.2 || 0.4|0.1 | 16|| 1122|30||38.4|11.5||47.7|14.3||10.3| 3.1|| 3.3| 0.9 || 0.2|0.06| 24|| 1722|35||40.1|14.0||35.0|12.2||23.6| 8.3|| 1.0| 0.3 || 0.1|0.03| 30|| 900|30||36.6|10.9||44.4|13.3||18.4| 5.5|| 0.1| 0.03|| 0.3|0.09| June 5|| 825|33||28.4| 9.4||49.3|16.2||20.0| 6.6|| 1.7| 0.6 || 0.4|0.1 | 12|| 1314|33||28.0| 9.3||49.0|16.2||20.0| 6.6|| 2.2| 0.7 || 0.8|0.3 | 19|| 917|37||32.4|11.9||52.3|19.3||14.5| 5.4|| 0.6| 0.3 || 0.2|0.07| 28|| 802|42||30.5|12.8||56.4|23.7||11.7| 4.9|| 0.7| 0.3 || 0.4|0.2 | July 2|| 1062|56||16.5| 9.2||57.1|31.9||25.6|10.3|| 1.2| 0.7 || 1.2|0.7 | 9|| 1245|51||17.6| 8.9||59.1|30.1||21.8|11.1|| 0.8| 0.4 || 0.8|0.4 | 16|| 974|69||17.5|12.0||66.4|45.8||15.7|10.8|| 0.2| 0.1 || 0.2|0.1 | 23|| 1156|58||21.7|12.6||67.2|38.9|| 9.5| 5.5|| 1.5| 0.9 || 0.2|0.1 | 30|| 802|54||20.2|10.7||65.4|34.6||12.8| 6.8|| 1.4| 0.7 || -- | -- | Aug. 6|| 910|52||21.7|11.3||67.3|34.9|| 9.7| 4.9|| 1.0| 0.5 || 0.3|0.2 | Sept. 6|| 815|51||23.0|11.7||65.3|33.5|| 9.8| 4.9|| 0.9| 0.5 || 0.4|0.2 | Oct. 5|| 625|62||26.4|16.3||64.4|39.9|| 8.5| 5.2|| 0.6| 0.4 || -- | -- | Nov. 4|| 800|58||22.5|13.0||66.4|38.5|| 9.6| 7.3|| 0.9| 0.5 || 0.5|0.2 | || | || | || | || | || | || | | 1889 || | || | || | || | || | || | | April 10|| 700|--||29.8| -- ||53.3| -- ||14.8| -- || 1.2| -- || 0.6| -- | June 6|| 900|71||28.2|20.0||50.1|35.6||12.9| 9.1|| 8.2| 5.8 || 0.6|0.4 | Aug. 1|| 670|62||30.6|18.9||44.2|27.4||15.2| 9.4|| 9.6| 5.9 || 0.4|0.2 | Dec. 4|| 731|63||36.0|22.0||38.3|24.1||11.3| 7.1||13.3| 8.7 || 0.6|0.4 | || | || | || | || | || | || | | 1890 || | || | || | || | || | || | | Feb. 2|| 622|51||32.3|16.5||30.1|15.3||11.1| 5.6||26.0|13.2 || 0.5|0.2 | April 30|| 500|48||36.5|17.5||24.5|11.7|| 9.4| 4.5||28.9|13.9 || 0.6|0.3 | ---------------------------------------------------------------------------