Improved detection of a staphylococcal infection by monomeric and protein A-purified polyclonal human immunoglobulin (original) (raw)

1993, European Journal of Nuclear Medicine

The present study was untertaken to compare the technetium-99m labelled non-specific polyclonal human immunoglobulin (Ig) with 99mTc-labelled monomeric human immunoglobulin (m-Ig), 99mTc-labelled, protein A-purified, human immunoglobulin (A-Ig) and 99mTc-labelled monomeric, protein A-purified, human immunoglobulin (mA-Ig) as tracer agents for the detection of a thigh infection with Staphylococcus aureus. In vitro the binding of the various tracer agents to bacteria at various intervals was determined. For the in vivo evaluation, mice were infected and received one of the various labelled proteins. Scintigrams were made 0.25, 1, 4 and 24 h later. All 99mTc-labelled Igs bound to bacteria in vitro: the percentages of binding for the m-Ig (from 1 h onwards) and A-Ig and mA-Ig (from 3 h onwards) were significantly higher than that for Ig. The in vivo target-tonon-target (T/NT) ratios were significantly higher from 4 h onwards for all purified Igs than for Ig. Protein A-purified Igs yielded higher T/NT ratios than m-Ig. Furthermore, the amount of activity in the liver was significantly lower 24 h after administration of m-Ig, A-Ig and mA-Ig than after administration of Ig. It is concluded that in this experimental infection 99mTc-labelled monomeric Ig localizes a staphylococcal thigh infection better and faster than 99mTc-labelled unpurified Ig. However, the accumulation obtained with protein A-purified Ig or protein Apurified monomeric Ig was the highest of all tracer agents tested.

Detection of a local staphylococcal infection in mice with technetium-99m-labeled polyclonal human immunoglobulin

Journal of nuclear medicine : official publication, Society of Nuclear Medicine, 1991

The purpose of this study was to investigate both the ability of 99mTc-labeled polyclonal human immunoglobulin (HIG) to localize an infection and the modes of action involved in this process. Mice, infected with Staphylococcus aureus ATCC 25923 in a thigh muscle, received HIG intravenously. Scintigrams were made 1, 4, and 24 hr later; subsequently the mice were killed and the activity in several organs and thighs was determined. The radiopharmaceutical demonstrated a time-dependent accumulation at the site of infection. It was found that vascular permeability or Fc binding alone could not account for the mode of action of HIG. Neither the origin of Ig (human versus murine) nor the total amount of protein (0.01-1.0 mg Ig per mouse) affected the target-to-background (T/B) ratios. Ratios were not different for leukocytopenic animals. A correlation (p less than 0.001) was demonstrated between the number of bacteria at the site of infection and the T/B ratio. This was also found after an...

Optimized localization of bacterial infections with technetium-99m labelled human immunoglobulin after protein charge selection

European Journal of Nuclear Medicine, 1994

To improve the scintigraphic detection of bacterial infections a protein charge-purified fraction of polyclonal human immunoglobulin was applied as a radiopharmaceutical. This purification was achieved by attaching the immunoglobulin to an anion-exchanger column and by obtaining the column-bound fraction with buffer. The binding to bacteria in vitro and the target to non-target ratios of an experimental thigh infection with Staphylococcus aureus or Klebsiella pneumoniae in mice were evaluated to compare the purified and the unpurified immunoglobulin. The percentage of binding to all gram-positive and gramnegative bacteria used in this study was significantly (P<0.03) higher for the purified than for the unpurified immunoglobulin. For the in vivo study, mice were infected in the thigh muscle with Staph. aureus or K. pneumoniae. After 18 h 0.1 mg of technetium-99m labelled polyclonal immunoglobulin or 99mTc-labelled protein charge-purified polyclonal human immunoglobulin was administered intravenously. At all time intervals the target (infected thighs) to non-target (noninfected thighs) ratios for both infections were significantly higher (P<0.03) for protein charge-purified polyclonal immunoglobulin than for unpurified polyclonal human immunoglobulin. Already within 1 h the infected tissues could be detected by the purified immunoglobulin. It is concluded that 99mTc-labelled protein charge-purified immunoglobulin localizes both a gram-positive and a gram-negative thigh infection more intensely and faster than 99mTc-labelled unpurified immunoglobulin.

Binding of99mTc-labelled polyclonal human immunoglobulin to bacteria as a mechanism for scintigraphic detection of infection

European Journal of Nuclear Medicine, 1991

The aim of the present study was to determine whether 99mTc-labelled polyclonal human immunoglobulin (99mTc-HIG) binds to bacteria in vitro as well as in vivo. In vitro, the binding of 99mTc-HIG to various gram-positive and gram-negative bacteria was determined. In vivo, mice were infected with Staphylococcus aureus Cowan I (protein A rich) or S. aureus EMS (protein A deficient) in a thigh muscle and then 99mTc-HIG or 99mTc-labelled human serum albumin (99mTc-HSA) was administered; scintigrams were made 1, 4, and 18 h later. In vitro binding of 99mTc-HIG to bacteria was higher for gram-positive than for gram-negative forms. A positive correlation was found between the protein A content and the degree of binding to S. aureus. This was also found in vivo. The accumulation of 99mTc-HIG at the site of infection was significantly (P< 0.01) higher than that of 99mTc-HSA, for both strains of S. aureus. It is concluded that vascular permeability cannot fully explain the accumulation of 99mTc-HIG at the site of infection and that binding of 99mTc-HIG to bacteria plays a role in this respect.

The measurement of staphylococcal protein A by ELISA in immunoglobulin preparations

Biologicals, 1991

Chicken antibodies were used to develop an ELISA for the quant~tatJon of parts-per-milhon levels of protein A in the purification of immunoglobuhns or 0mmunoglobuhn-like molecules Quant~tat~on of prote~n A ~n the presence of excess human or murlne ~mmunoglobuhns ~n this assay was compared with that obtained ~n ELISAs developed w~th rabbit antibodies specific either to prote~n A or to other molecules Experiments demonstrate that protein A is bound to the ~mmunoglobuhns being punfied and that this binding affects subsequent recognition by the antibodies used for the assay Because of these effects and because fragments of protein A might not be detected in assays which rely on Fc binding of protein A, chicken antibodies that bind protein A specifically are an advantage for the quantitat~on of th~s protein by ELISA In addition, comparison of the effect of different types of immunoglobuhns on the proteMn A standard curve suggests that alternatives to including the immunoglobulin under punficataon with the standards can be ut~lMzed Materials and methods Reagents Chicken antibodies to protein A, both unconjugated and conjugated to horseradish peroxidase

A kit of human polyclonal IgG for the diagnosis of infectious processes

Journal of Radioanalytical and Nuclear Chemistry, 1999

The aim of this work was to obtain a freeze-dried kit for direct 99mTc-labeling of human polyclonal IgG. The labeling procedure was carried out by Schwarz's method. The best yields of 99mTc-IgG were obtained by using sodium pyrophosphate decahydrate as a weak chelating agent. Performed tests showed the stability of the radiopharmaceutical up to 24 hours. Plasma clearance in rats was fitted to a biexponential curve with Tl/2u = (0.1+ 0.9) h and Ti/2f ~ = (10-Z-_3) h. The organs with higher uptake of radiopharmaceutical were lung, kidneys and blood. In a rabbit model the abscess target/background ratio was 3-6 according to time of the scintigraphic images. Thirty patients with musculoskeletal infection were studied. Twenty-one lesions were detected and confirmed by culture/biopsy. *

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